Research Area Center for Molecular Biosciences (CMBI)

scientific report | 2016/2017

Scientific Coordinators Bert Hobmayer, Ronald Micura, Jörg Striessnig 2  Imprint  3

The Research Area Center for Molecular Biosciences Innsbruck (CMBI) – a life science network in western Austria

In our biannual report, the Center for Molecular Biosciences of Innsbruck University (CMBI) presents its recent scientific achievements, new developments in ongoing research projects and success stories of its faculty members, especially of young researchers. Molecular biosciences represent one of the most exciting fields of modern research among the natural sciences. They bridge the gap between single molecules and the complex functions in living organisms under normal conditions and in disease. Minor changes in bioactive molecules such as DNA, RNA and proteins affect and change the properties of cells, microorganisms, and plants. Advances in technologies including microscopic imaging, new generation sequencing applications and techniques to analyze molecular structures result in an explosion of information and understanding of biological systems primarily oriented to improve human health. The CMBI aims at providing a platform for this extremely rapidly developing research field by taking advantage of the visibility and expertise of the CMBI’s internationally competitive groups to strengthen interdisciplinary research activities.

The CMBI currently consists of 21 research teams originating from the faculties of Chemistry and Pharmacy, of Biology, and of Mathematics, Informatics and Physics, and their activities focus on research and teaching. CMBI members contribute to the FWF special research program SFB-F44 “Cell Signaling in Chronic CNS Disorders”, which is currently in its second funding period, and to several FWF-funded doctoral programs, all in collaboration with the Medical University of Innsbruck. Notably, several member labs have now initiated a first CMBI-embedded university PhD program under the topic “Ageing and Regeneration”. Furthermore, CMBI members were able to successfully compete in the recent FFG call for university infrastructure and now establish new facilities for Nuclear Magnetic Resonance (NMR) spectroscopy. These will significantly advance biomolecular structural analysis in the Western-Austrian area. In order to cross-link with the Tyrolean Life Science community and to foster career perspectives of young scientist, the CMBI is a co-organizer of the internationally visible Innsbruck Life Science meetings, and it has started to coordinate its activities as a core research unit within the newly established initiative “Tyrolean Health and Life Science Cluster”.

In spite of these achievements, the CMBI faces challenges of general and also local nature. Rapid imprint | technological progress in the Life Sciences requires ongoing investments into essential state-of the art infrastructure. Ever-increasing demand for big data sets needs development of adequate computing units and bioinformatics expertise. More than half of the 21 research teams are located Content: Bert Hobmayer, Professor of Zoology Ronald Micura, Professor of Organic Chemistry together with Life Science research departments of the Innsbruck Medical University in the recently Jörg Striessnig, Professor of Pharmacology built Center for Chemistry and Biomedicine (CCB) building at the university’s central campus. Nine

Layout: Stephanie Brejla, Gabi Reiter of those, however, are spatially fragmented at the Technics Campus, the Department of Botany, and the Research Department for Biomedical Aging Research. Bringing together these research Contact: Research Area Center for Molecular Biosciences (CMBI) units within a single research campus would strongly stimulate scientific interactions and facilitate

the efficient use of expensive infrastructure. Long-term strategies towards this goal are needed to University of Innsbruck Innrain 80-82a enable competition with a large number of Life Science Research centers rapidly emerging across A-6020 Innsbruck Europe. All these issues require coordinated efforts at academic and political levels. Tel: +43 512 507-57501 Cover figure: Fax: +43 512 507-57599 Induced neural stem cells (iNSCs) being reprogrammed from human skin fibroblasts by the team of [email protected] CMBI group leader Frank Edenhofer. Colonies of proliferating iNSCs start to differentiate into neurons The CMBI coordinators: www.uibk.ac.at/cmbi in the periphery. Differentiating neurons exhibit characteristic outgrowth of TUJ1-positive neurites (green) that reach out to contact other neurons to build a functional network. Bert Hobmayer Jörg Striessnig Ronald Micura © 2018 by CMBI 4  Contents  5

Overview 6 CMBI - news CMBI - specials 10 Research Summary Signal transduction in cellular growth control and 18 carcinogenesis Advanced chromatographic and spectroscopic analytical 20 tools in natural product analysis & molecular biology Characterization of RNA, proteins, and their noncovalent 22 complexes by mass spectrometry Gastropod metallothioneins in evolution: New rules for an 24 old Radiation damage in biological compounds induced by low 26 energy electrons contents | Regenerative potential of reprogrammed neural stem cells 28 Immuno-gerontology 30 Development of selectively acting antitumor drugs 32 Developmental biology and bioadhesion in basal 34 model systems Molecular and cell biology of human aging 36 Plant biochemistry and metabolism 38 Chemistry, chemical and structural biology of the pigments 40 of life Protein dynamics and biomolecular recognition 42 Developmental biology 44 Synthesis, structure, and function of non-coding RNAs 46 Cell physiology and gene regulation 48 Structure-functional activity relationship investigations on 50 ligands interacting with opioid receptors Targeted proteolysis in human diseases and its impact on 52 drug development and production Cell signaling in chronic CNS disorders 54 Pharmacognosy – combining traditional knowledge with 56 innovation Biomolecular NMR spectroscopy 58 Publications 60 CMBI - careers of young researchers 92 Awards & Honors for CMBI scientists 96 CMBI Meetings and Seminar Series 99 6  Overview Overview  7

>> CMBI facts The Research Area Center for Molecular Biosciences (CMBI) at the University of Innsbruck is an integrative and multidisciplinary research and teaching institution. The mission of the CMBI is to advance studies on the structure, function, and interaction of biological macromolecules and low molecular weight compounds relevant for cellular growth, metabolism, and development. The research activities in the CMBI take advantage of existing research strength in different fields and have strongly promoted interdisciplinary research activities in five major fields of biomolecular sciences.

Basic and applied biomolecular research fields at the CMBI • Structure, dynamics and interactions of biologically important molecules • Molecular basis of physiological and pathophysiological processes • Metabolites, natural and synthetic compounds that modulate important biological processes • Cell-to-cell communication and cellular function • Development, regeneration and aging of whole organisms

Twenty-one research groups from the faculty of Chemistry and Pharmacy, the faculty of Biology, and the faculty of Mathematics, Informatics & CMBI members received scientific awards and prizes over the last two Physics are members of the CMBI. years thus documenting their successful research activities also to the scientific community and general public. Among those are the Bruker research award 2017 and the Research Award of the Stiftung Südtiroler Sparkasse 2017 to Hermann Stuppner, the Edmund Optics Educational CMBI members Areas of expertise Award Europe 2017 to Torsten Schwerte, and the Würdigungspreis des Chemistry Bundesministeriums für Wissenschaft, Wirtschaft und Forschung 2017 K. Bister, M. Hartl, E. Stefan biochemistry, molecular genetics G. Bonn, C. Huck bioanalytics to Anita Siller. Ilse Kranner was elected as President of the Austrian K. Breuker biomolecular mass spectrometry Society of Plant Biology (ATSPB). Furthermore, several CMBI members B. Kräutler, T. Müller structural biology, natural products chemistry are members of the Austrian Academy of Sciences (Kathrin Breuker, K. Liedl, D. Schuster theoretical chemistry, computer-aided molecular design Ronald Micura, Bernhard Kräutler, Jörg Striessnig) and of the German R. Micura chemical biology of nucleic acids Academy of Sciences, Leopoldina (Bernhard Kräutler, Jörg Striessnig). R. Schneider, B. Auer molecular biology, biotechnology The activities of the CMBI are currently coordinated by Bert Hobmayer M. Tollinger, C. Kreutz biomolecular NMR spectroscopy (head), Jörg Striessnig and Ronald Micura. Pharmacy R. Gust medicinal chemistry, drug design In the years 2013 - 2017 the CMBI member labs published 737 papers H. Schmidhammer, M. Spetea pharmaceutical chemistry, drug design in peer reviewed journals. This includes 28 publications in the world J. Striessnig, A. Koschak, N. Singewald cell biology, neuropharmacology leading journals Nature, Nature Structural Biology, Nature Genetics, H. Stuppner, M. Ganzera pharmaceutical biology, phytochemistry Nature Cell Biology, Nature Chemical Biology, Nature Neuroscience,

members | Biology Nature Communications, Nature Methods, Proceedings of the R. Dallinger cell physiology, ecotoxicology National Academy of Sciences of the United States of America, and F. Edenhofer stem cell biology Scientific Reports, 2 papers in Cell and 38 papers in top journals of B. Grubeck-Loebenstein immuno-gerontology chemistry and physics, including Journal of the American Chemical B. Hobmayer, P. Ladurner cell and developmental biology P. Jansen-Dürr cell biology, molecular biology Society, Chemical Society Reviews, Angewandte Chemie and EMBO I. Kranner plant physiology and biochemistry J. The total amount of outside grant support of the last three years D. Meyer, P. Aanstad developmental biology amounts to about 9,4 Mio. €. Modern infrastructure obtained through B. Pelster, A. Sandbichler, T. Schwerte cell biology, cell physiology special governmental funding for research equipment significantly Physics strengthens research in structural and cell biology, bioanalytics and S. Denifl biophysics, radiation physics biophysics at the CMBI. 8   9

The specific research topics of the 21 CMBI member labs are listed below.

Research topics

• Protein and nucleic acid structure, folding, and dissociation in the gas phase (Breuker) • Proteomics, metabolomics, phytomics (Bonn, Huck) • Inelastic interaction of low energy electrons with molecules of biological relevance (Denifl) • Development of theoretical and computational methods describing molecular interac- tions in chemical and biological systems (Liedl, Schuster) • Biomolecular NMR spectroscopy (Tollinger, Kreutz)

• Synthesis, structure, and function of chemically modified RNA (Micura) topics | • Regulation of cell function by protein modification (Auer, Schneider) • Oncogenic transcription factors and their cellular targets (Bister, Hartl, Stefan) • Natural products chemistry, chemical and structural biology of the pigments of life (Kräutler, Müller)

• Bioactive natural products from the plant kingdom (Stuppner, Ganzera) • Development of potential drugs interacting with opioid receptors (Schmidhammer, Spetea) • Development of selectively acting antitumor drugs (Gust)

• Ion channels as new drug targets and the neuropathological basis of anxiety disorders (Striessnig, Koschak, Singewald) • Cell ion and volume homeostasis and metabolic activity (Pelster, Sandbichler, Schwerte) • Trace element homeostasis in animal cells (Dallinger)

• Molecular and genetic control of vertebrate development (Meyer, Aanstad) • Stem cell differentiation, regeneration and bioadhesion of basal Metazoa (Hobmayer, Ladurner) • Stem cell biology, cellular reprogramming & regeneration (Edenhofer)

• The aging T cell repertoire and its impact on vaccinations of the elderly (Grubeck-Loe- benstein) • Biology of aging, mitochondrial physiology (Jansen-Dürr) • Stress metabolites and signalling pathways in plants (Kranner) 10  CMBI news CMBI news  11

>> ELUCIDATION OF RIBOZYME STRUCTURES AND MECHANISMS CMBI - specials

Three years ago, four new classes of ribozymes were discovered. Like , these RNA accelerate chemical reactions in the cell. A research group led by Ronald Micura and researchers in China around Aiming Ren have now elucidated the structure of the so-called Twister-Sister ribozyme. The sequence design and the syntheses for the investigations were done in Innsbruck, the crystallization and X-ray structure analysis was undertaken by the scientists at Zhejiang Structural model of ribozyme function University in Hangzhou. Moreover, the Micura group also investigated the Pistol ribozyme and elucidated its chemical mechanism. The picture: Otto Peter functional analysis was laborious, because the chemists had to synthesize numerous new variants of this RNA, each of which only a single atom was exchanged. Only then was it possible to check which >> NEW NMR-SPECTROMETERS The structural dynamics of nucleic acids and their interactions with of the modified ribozymes is still functional. From this, it was possible proteins or small molecule compounds has already been successfully to deduce the positions responsible for reaction catalysis in the RNA researched at the University of Innsbruck in several working groups. structure. With the acquisition of new nuclear magnetic resonance (NMR) spectrometers with frequencies of 400 and 700 MHz, this research Zheng L et al. Structure-based insights into self-cleavage by a four-way junctional twister-sister ribozyme. Nature Communications 8, 1180 (2017). focus will be further strengthened. With their application to the 1st Neuner S et al. Atom-Specific Mutagenesis Reveals Structural and Catalytic Roles for an Active-Site F&E Infrastructure Call (2016) of the Austrian Research Promotion Adenosine and Hydrated Mg2+ in Pistol Ribozymes. Angew. Chem. Int. Ed. 56, 15954–15958 (2017). Agency FFG, the Innsbruck scientists around Ronald Micura, Christoph Kreutz, Martin Tollinger, Kathrin Breuker, Ronald Gust, and Hermann Stuppner prevailed in a very tough Austria-wide competition. “The >> SEEING THE INVISIBLE – RNA AND DNA EXCITED STATES new devices expand the possibilities in the area of structural chemistry and they complement the existing spectrometers, which have been Using nuclear magnetic resonance (NMR) spectroscopy, unexpected fully utilized for several years”, says a delighted Ronald Micura. high-energy excited states of nucleic acids could be detected and The new infrastructure will further enhance the international visibility characterized. These transiently existing conformations are present of the various fields at the Center for Molecular Biosciences. Several at < 10%, but they represent important functional intermediates research groups are concerned here with the structure and folding in cellular processes. These states were addressed via novel NMR of RNA and their interaction with low molecular weight compounds. spectroscopic methods. It could be shown, that a small, undesired Complex dynamic folding mechanisms are involved, which can be modification of DNA, a methylation, induces highly localized elucidated by means of NMR spectroscopy. Another focus is on the transiently populated structural alteration in base pairing. These study of RNA-modifying enzymes and proteins that can trigger allergies Models of high-energy excited states of nucleic acids high energy states very likely serve as a beacon for repair enzymes or are relevant to inflammatory signaling pathways, and in the search and are therefore very important for the preservation of genetic for plant-derived natural products that can exert antibacterial effects information. In contrast, naturally occurring RNA methylations lead via RNA target molecules. to drastic structural changes and expand the functional properties “The wealth of information that can be made accessible by NMR of RNA. The different behavior of DNA and RNA could also explain spectroscopy is an ideal complement to the ‘static’ crystallographic why nature stores the genetic information in the double helix of DNA images of biomolecular systems, and it is poised to become the and not in the evolutionarily much older RNA. The work was done method of choice for studying the dynamic interaction networks of by international cooperation with working groups at Duke University biomolecules for understanding in cells”, says Ronald Micura. Based (USA), Max F. Perutz Laboratories in Vienna and University College on the new infrastructure, the research groups of Robert Konrat from London (UK). the Max F. Perutz Laboratories in Vienna, Fatima Ferreira-Briza at

With a new infrastructure worth two million Euros, interdisci- the University of Salzburg and Michael Oberhuber at the Laimburg Juen MA et al. Excited States of Nucleic Acids Probed by Proton Relaxation Dispersion NMR plinary research on RNA at the University of Innsbruck will be Spectroscopy. Angew. Chem. Int. Ed. 55, 12008 (2016). Research Center in South Tyrol, who are involved in comprehensive further strengthened. Since November 2017, two new NMR Zhou H et al. m1A and m1G disrupt A-RNA structure through the intrinsic instability of Hoogsteen spectrometers are available to scientists. scientific cooperation, will also benefit. base pairs. Nature Structural & Molecular Biology 23, 803–810 (2016). 12  CMBI news CMBI news  13

CMBI - specials

>> MOLECULAR BASIS OF CANCER AND OTHER CHRONIC DISEASES >> GAP JUNCTIONAL INTERCELLULAR COMMUNICATION ESSENTIAL FOR ENDODERM FORMATION Klaus Bister from the Department of Biochemistry and Eric Hunter

from the Emory University (Atlanta) have served as editors for a special An international research team including the group of Frank Edenhofer volume - Viruses, Genes, and Cancer - of the book series Current Topics investigated the role of gap junctional intercellular communication in Microbiology and Immunology. The editors were able to engage (GJIC) by generating Connexin43/Connexin45-double deficient mouse eminent researchers from the cancer field as contributors for this embryonic stem cells. These Connexin double-knock-out cells show volume, including the Nobel laureates Mike Bishop (San Francisco), almost complete breakdown of GJIC and a block of differentiation Harold Varmus (New York), and Harald zur Hausen (Heidelberg). The in embryoid bodies. However, pluripotency marker expression and chapters focus on virus-host cell interactions, cellular genes acquired proliferation are unaffected. They fail to form primitive endoderm or modulated by viruses, the pathological effects of these interactions, in embryoid body cultures, representing the inductive key step of Development of embryonic stem cells of the inner cell mass and therapeutic interventions. Several articles specifically address the into endodermal and ectodermal tissue layers gastrulation-like events. At a molecular level, GJIC-incompetent role of viruses and genes – such as oncogenes, proto-oncogenes, or embryonic stem cells exhibit significantly less activated NFATc3 in tumor suppressor genes – in the etiology of human cancer. Oncogenic cellular nuclei than control cells suggesting that Connexin-mediated signaling by PI3 kinase, mTOR, Akt, or the major cancer drivers MYC communication is needed for synchronized NFAT activation to induce and RAF, and the role of tumor suppressors like p53, are discussed orchestrated primitive endoderm formation. in detail. The volume also explores the emerging role of noncoding Wörsdörfer P et al. Abrogation of gap junctional communication in ES cells results in a disruption of RNAs such as microRNAs in tumorigenesis and cancer therapeutics. primitive endoderm formation in embryoid bodies. Stem Cells 35, 859-871 (2017)

Hunter E, Bister K (eds). Viruses, Genes, and Cancer - Current Topics in Microbiology and Immunology, Vol. 407. Springer International Publishing (2017). Stefan E, Bister K. MYC and RAF: Key effectors in cellular signaling and major drivers in human cancer. Curr. Top. Microbiol. Immunol. 407, 117-151 (2017). >>NEW ROLE FOR MYC IN STEM CELL DORMANCY AND SELF RENEWAL >> „INNSBRUCK” MODEL ORGANISM: TRANSGENESIS ESTABLISHED

Researchers of the German Cancer Research Center and the team of The free-living flatworm Macrostomum lignano is a suitable model Frank Edenhofer at the Institute of Molecular Biology have identified system for regeneration, reproduction, evolution and bio-adhesion that Myc depletion in mouse embryonic stem cells results in a research. However, a major cornerstone of research methods – the reversible biosynthetic dormancy and proliferation arrest. Likewise, ability to generate transgenic animals – was missing for Macrostomum Myc inhibition in mouse blastocysts induces dormancy mimicking and the whole flatworm community. In cooperation with colleagues hormonally controlled diapause without affecting the pluripotency from the European Institute for the Biology of Ageing (Groningen, capacity, suggesting the importance of Myc regulation in controlling NL), the Ladurner group published the generation of stable entry and exit from stem cell dormancy during development. transgenic Macrostomum lines expressing fluorescent proteins under Furthermore, Myc-depleted stem cells and diapause embryos do several tissue-specific promoters. The reported method will permit exhibit similar expression signatures. Thus, dormancy is a transient sophisticated research approaches including gene overexpression, stem cell state, molecularly defined by low levels of Myc expression, dissection of gene regulatory elements, real-time imaging and lineage where the most primitive cells within the stem cell compartment can tracing. It provides a technological platform for harvesting the power Working model for Myc-induced dormancy in stem cells be stored as a transiently quiescent reservoir to preserve genomic of Macrostomum as an experimental model organism for flatworm integrity and be protected from microenvironmental hazards and

Various transgenic Macrostomum lines expressing biology and biomedical research. replicative stress. fluorescent proteins under tissue-specific promoters Wudarski J et al. Efficient transgenesis and annotated genome sequence of the regenerative Scognamiglio R et al. Myc depletion induces a pluripotent dormant state mimicking diapause. Cell flatworm model Macrostomum lignano. Nature Communications 8, 2120 (2017). 164, 668–680 (2016). 14  CMBI news CMBI news  15

CMBI - specials

>> DISCOVERY OF DIFLAPOLIN AS AN ANTI-INFLAMMATORY SUBSTANCE >> CHLOROPHYLL AND VITAMIN B12 - TWO ‘PIGMENTS OF LIFE’

Using a combination of pharmacophore models for soluble epoxide Recent research of Bernhard Kräutler and his group brought major and 5-lipoxygenase activating protein, Daniela Schuster further discoveries concerning the biological role of the seasonal and her co-workers discovered and participated in a detailed disappearance of chlorophyll, as well as on the metabolic function and molecular logic of vitamin B . characterization of a potent dual inhibitor targeting both proteins 12 in sub-micromolar concentrations. The substance, which was The abundant and highly visual green leaf pigment chlorophyll is named Diflapolin, selectively inhibited leukotriene formation and broken down in plants by strictly regulated biological processes epoxyeicosatrienoic acid degradation, and was not active against furnishing ‘phyllobilins’ in an amount of 1000 million tons each other targets from the arachidonic acid cascade. Importantly, it year on Earth. In contrast to all earlier views, phyllobilins are now showed anti-inflammatory activity in vivo in mice. A patent was filed presumed to have physiological functions that are still enigmatic, on this and structurally related compounds, and they are currently but possibly similar to those of related heme catabolites, such as Daniela Schuster (left) and Veronika Temml (right) optimized towards more favorable properties in collaboration with the yellow bilirubin. Attractive roles of phyllobilins in plants could Prof. Matuszczak and Mag. Vieider. concern photo-regulation, as well as pathogen defense. We have studied yellow phyllobilins recently and discovered their behavior as Phyllobilin structure and autumn leaves with disappearing Temml V et al. Discovery of the first dual inhibitor of the 5-lipoxygenase-activating protein and chlorophyll medium responsive photo-switches, a property that hints at a possible soluble epoxide hydrolase using pharmacophore-based virtual screening. Scientific Reports 7, 42751 (2017). role in photo-regulation. Garscha U et al. Pharmacological profile and efficiency in vivo of diflapolin, the first dual inhibitor In order to understand better some of the proposed enigmatic of 5-lipoxygenase-activating protein and soluble epoxide hydrolase. Scientific Reports 7, 79398 (2017). (‘moonlighting’) activities of the B12-cofactors, we have recently

suggested to study effects of ‘antivitamins B12’, structural analogs

of vitamin B12 that cannot function as the vitamin does. Thus, these types of ‘wolf in a sheep’s clothes’ inhibit the canonical enzymatic

>>MICRO-RNA MIR-144 AND ANXIETY DISORDER B12-functions, enabling researchers to determine the intact structures of key enzymes. In this context, a synthetic methodology to the

particularly intriguing group of analogs of vitamins B12 has been Effective long-term treatment for fear, trauma and anxiety-related recently opened up, in which the B12’s own cobalt center is replaced disorders is a continuing challenge. One emerging new treatment by another transition metal, such as the homologue rhodium. This strategy is combining exposure-based cognitive behavioural therapy extremely difficult chemical task was achieved in collaboration with extinction-facilitating drugs. We, in collaboration with national with biologists in England, providing the long awaited road to and international partners identified a specific microRNA, mir-144, fundamentally new B12-analogues and a new group of excellent that is critically involved in orchestrating the rescue of impaired inhibitors. extinction learning. Its expression was shown to be decreased in Ruetz et al. Antivitamin B inhibition of the human B -processing enzyme CblC: Crystal structure extinction-impaired mice and upregulated upon extinction learning 12 12 of an inactive ternary complex with glutathione as the cosubstrate. Angew. Chem Intl. Ed. 56, 7387 Nicolas Singewald and his team in extinction-competent mice. We identified an important locus of (2017). Li et al. Chlorophyll-derived yellow phyllobilins of higher plants as medium-responsive chiral action, since overexpression of mir-144 in the amygdala was sufficient photoswitches. Angew. Chem Intl. Ed. 55, 15760 (2016). to fully rescue the extinction learning impairment. Since specific Widner et al. Total synthesis, structure, and biological activity of adenosylrhodibalamin, the non- natural rhodium homologue of coenzyme B12. Angew. Chem Intl. Ed. 55, 11281 (2016). drugs have been shown previously to increase mir-144 expression, this route will be followed to develop drug-based strategies to boost and normalize aberrant fear-inhibitory learning, which is a common deficiency observed in human anxiety disorders.

Murphy CP et al. MicroRNA-mediated rescue of fear extinction memory by miR-144-3p in extinction- impaired mice. Biol. Psychiatry 81, 979-989 (2016). 16  CMBI news CMBI news  17

CMBI - specials

>> ARTEMISININS AS POTENTIAL COMPOUNDS IN DIABETES THERAPY? >> FWF-FUNDED DOCTORAL DOCFUNDS PROGRAM „CAVX“ STARTS AT THE INNSBRUCK UNIVERSITIES WITH CMBI PARTICIPATION Researchers from the Institute of Molecular Biology cooperated with Stefan Kubicek, Group Leader at the CeMM (Vienna), to demonstrate The CavX project has been established as a doctoral program with in vivo the beta cell-promoting effects of artemisinins, compounds a cell- and neurobiological focus on cellular signaling through ion which were discovered through screening efforts in cell lines to steer channels. The research groups of Alexandra Koschak, Petronel Tuluc alpha cells towards a beta cell fate. In work done by the group of and Jörg Striessnig are the CMBI project leaders in the newly FWF- Dirk Meyer, zebrafish with severely reduced beta cells were treated funded doc.funds project CavX together with colleagues from the with artemisinins. This resulted in increased beta cell numbers Medical University (Marta Campiglio, Bernhard E. Flucher and Gerald and normalized glucose levels. Chemicals which enhance beta cell Obermair, who also serves as a CavX coordinator). This program production have potential as therapies for diabetes, in which these provides a special platform for the training of graduate students in critical cells are lost. Follow up studies have been aimed to more cellular signaling in electrically excitable cells. The FWF doc.funds precisely define the effects of artemisinins. initiative was started only recently and has been awarded for the

Li J et al. Artemisinins target GABAa receptor signaling and impair α cell identity. Cell 168, 86-100 first time in fall 2017. 45 proposals were submitted and from those (2017). an international multidisciplinary jury selected 16 for a final hearing. Seven of those were finally recommended for funding. The application process of the CavX program was strongly supported and guided by the vice-rectors for research of the two participating universities.

>> START OF THE INTERREG V-A ITALIA-ÖSTERREICH PROJECT „APPLECARE“ >> NEWLY IDENTIFIED STRESS PROTEIN PROTECTS SNAILS FUNDED BY THE EUROPEAN REGIONAL DEVELOPMENT FUND

Snails are model organisms for research on metal stress and In a transnational approach combining research competences in detoxification of heavy metals. As other animals, they use structural chemistry, medicine and molecular biology, the molecular Metallothionein proteins to bind and neutralize heavy metals such details of apple allergy are studied and potential apple varieties for as cadmium. An international research team organized by Reinhard therapy of birch pollen allergy are identified. Roughly 20% of the mid- Dallinger including collaborators in Zurich and Barcelona succeeded in European population are affected by birch pollen allergy, and more isolating and characterizing an unexpected form of Metallothionein than 70% of them subsequently develop allergic reactions to apples protein from a marine snail, which exhibits three instead of two metal- due to cross-reactivity of IgE antibodies between homologous proteins binding motifs. Thereby, one of these proteins is able to bind nine in birch pollen and apples. This immunological cross-reaction may Cadmium ions, much more than other Metallothioneins, resulting in present an opportunity to use apples in a controlled and established a much higher detoxification efficiency and higher stress resistance of The “applecare” team at the CMBI: R. Eidelpes, L. Ahammer, dosage to cure birch pollen allergy. Using NMR spectroscopy, the theses snails. Identification of this new protein structure is a result of J. Unterhauser and M. Tollinger (left to right) three-dimensional structures of different isoforms of the major apple a FWF DACH project on the evolution of the Metallothionein protein allergen, the protein Mal d 1, and the birch pollen allergen Bet v 1 are family funded together with the Swiss National Science Foundation compared in order to identify those apple varieties ideal for successful SNSF. long-term therapy of birch pollen allergy by apple consumption. Within the framework of the project “applecare”, complementary Baumann C et al. Structural adaptation of a protein to increased metal stress: NMR structure of a marine snail Metallothionein with an additional domain. Angew. Chem. Int. Ed. 56, 4617-4622 research competences in Tyrol and South Tyrol are pooled, including (2017). the Laimburg Research Centre, Bolzano Hospital, Innsbruck Medical Commented on by: Fahrenkamp-Uppenbrink L. Heavy metals? No problem for this snail. Science 356, 150-151 (2017). University and the University of Innsbruck. 18  Research summary Research summary  19

Signal transduction in cellular growth control and carcinogenesis

Klaus Bister, Markus Hartl, and Eduard Stefan

>> Department of Biochemistry

Myc is regulated by several signaling cascades implicated in development and oncogenesis. The Wnt/β-catenin/Tcf axis drives cell differentiation and organismal patterning. Disturbances of this pathway - like mutation of the APC tumor suppressor in colon cancer - leads to tumorigenic processes including transcriptional deregulation of the c‑myc protooncogene. The early diploblastic cnidarian Hydra has >> Goal: Molecular mechanisms of cellular growth control and two myc genes (myc1, myc2), with myc2 being closer related to c‑myc. carcinogenesis Wnt/β-catenin signaling has crucial functions in cell differentiation and development in Hydra. In collaboration with the Hobmayer group, Figure 1: Schematic diagram of basic interconnections between Myc and Ca2+/CaM signaling. The binding reactions >> Background: Research in our lab is focused on the analysis of signal we found that activation of Wnt/β-catenin signaling in Hydra leads and equilibria involved are indicated by black dashed arrows, transduction pathways, mediated by plasma membrane receptors, to downregulation of myc1 but not of myc2. Mapping and analysis cytoplasmic proteolytic processing or nuclear transport of Myc is depicted by red or green arrows, respectively. Specific second messengers, cytoplasmic kinases, and transcriptional regulators. of the myc1 and myc2 promoter regions revealed the presence of cleavage to Myc-nick requires the action of Ca2+-dependent Both physiological and pathogenic forms of these pathways are consensus binding sites for the transcription factor Tcf. In vivo binding proteases (red dashed arrow). (Raffeiner et al., Oncotarget 2017) investigated, with specific focus on their role in carcinogenesis. of Hydra Tcf to both promoter regions was demonstrated by chromatin immunoprecipitation. Reporter gene assays showed that the myc1 >> Research Highlights and Outlook: The bHLH-LZ (basic region/ promoter is repressed by ectopic Hydra β‑catenin/Tcf, whereas the myc2 helix-loop-helix/leucine zipper) oncoprotein Myc and the bHLH-LZ promoter was not affected. The identification of the Wnt/β-catenin/ protein Max form a binary transcription factor complex controlling Tcf axis as a regulatory pathway of Hydra myc genes indicates that fundamental cellular processes. Deregulated Myc expression leads to principal links in the Wnt signaling network have emerged very early in neoplastic transformation and is a hallmark of most human cancers. Myc metazoan evolution. transcription factor activities are controlled by defined protein-protein At the plasma membrane an array of more than 800 G protein-coupled interactions (PPI). We have recently obtained evidence for a second receptors (GPCRs) receive, convert, amplify, and transmit incoming Figure 3: PPI network emanating from PKA. Following phos- messenger controlled physical interaction between the Ca2+ sensor phoproteomic analyses using LC-MS/MS, a static PPI network signals. Activated GPCRs team-up with intracellular scaffolding proteins calmodulin (CaM) and all Myc variants (v-Myc, c‑Myc, N‑Myc, and L-Myc). of the confident set of PKA interacting proteins (endogenous to compartmentalize signal transmission. Scaffolds, such as β-arrestin complexes isolated from osteosarcoma cells) was generated. The predominantly cytoplasmic Myc:CaM interaction is Ca2+-dependent, (Bachmann*, Mayrhofer* et al., PNAS 2016). and A-kinase anchoring proteins (AKAPs), function as a physical nexus and the maps to the conserved bHLH domain of Myc. between receptors and molecular switches. Typically, these receptor- Ca2+-loaded CaM binds the monomeric and intrinsically disordered Myc bound AKAPs recruit protein kinase A (PKA) to assemble dedicated protein with high affinity, whereas Myc:Max heterodimers show less, polyvalent signaling complexes that are spatially and temporally >> Research Grants and Max homodimers no affinity for CaM. NMR spectroscopic analyses confined. We showed that the orphan GPCR, Gpr161, is a PKA substrate FWF: P23652 (2011-2016), P27606 (2015-2018), P30441 corroborate the biochemical results on the Myc:CaM interaction and (2017-2021), SFB-F44 associated member (2015-2016), and also has an AKAP motif embedded in its C-terminal tail. Our results Figure 2: Schematic depiction of the canonical Wnt signalling TWF: UNI-0404/1525 (2014-2016); TWF: UNI-0404/1990 pathway. Wnt binds to the Frizzled (FZD) receptor leading confirm the interaction site mapping. Cell-based reporter analyses and suggest that Gpr161, by directly recruiting type I PKA holoenzymes (2017), Alfonso M. Escudero fellowship (2015-2016) to disruption of the APC/GSK3b complex which normally cell transformation assays suggest that increasing CaM levels enhance to the receptor, creates a cAMP-sensing signalosome. Furthermore, degrades b‑catenin (b-Cat). Accumulated b-Cat translocates >> Coworkers into the nucleus where it binds to the Tcf transcription factor Myc transcriptional and oncogenic activities. Our results point to a V. Bachmann, P. Raffeiner, O. Torres-Quesada (postdocs); we propose that Gpr161 plays a role in recruiting isoform-specific leading to transcriptional activation of Wnt signalling targets possible involvement of Ca2+ sensing CaM in the fine-tuning of Myc M. Bucher, F. Enzler, A. Feichtner, J. Mayrhofer, R. Röck, A A PKA complexes to primary cilia. Currently we analyze and perturb the via Tcf binding elements (TBE, 5’‑CCTTTG /T /T-3’). (Gufler et C. Schneider, B. Texler (master and Ph.D. students); S. al., Dev. Biol. 2018, Epub 2017; Hartl et al., unpublished). function. Beiler, K. Puglisi, A. Raffeiner, A. Reintjes (technicians) receptor-effector relationship in the primary cilium. 20  Research summary Research summary  21

Advanced chromatographic and spectroscopic analytical tools in natural product analysis & molecular biology

Christian Huck and Günther Bonn

>> Department of Analytical Chemistry and Radiochemistry

>> Goal: Novel chromatographic and vibrational spectroscopic tools with >> Research Highlights and Outlook: The realization of polymeric enhanced efficiency, selectivity and sensitivity, are developed in order to monolithic sorbents with retention mechanisms based on more than get detailed analytical information upon the composition, origin and/or one type of interaction was realized and this type is referred to as mixed species of samples in the fields of phytomics, metabolomics, proteomics mode stationary phases. Fast gradient separations allowed the analysis and foodomics. of alkyl benzenes and parabens within a few minutes. Compared to silica based particles, the polymeric stationary phase provides pH >> Background: The development of novel monolithic stationary phases stability over the complete pH range and its preparation as monolith for the separation of small molecules was highly successfully continued. in capillary successfully shows several advantages compared to packed

Those stationary phases have been designed for either sample Figure 2: Strategy for the development of an on-line quality columns. preparation by solid phase extraction (SPE) or analytical separations by assurance procedure of medicinal plants Polymeric mixed-mode stationary phases are of high importance high performance liquid chromatography (HPLC). Thereby, the monolithic regarding their use as small SPE columns in sample preparation format offers several advantages, including superior efficiency, high approaches. For that polymeric mixed-mode sorbents were based on permeability, mechanical robustness, and extended useful life-circle. an imidazole network and feature auspicious characteristics including New methodologies for solid-phase extraction (SPE) based on polymeric, a high polarity, an aromatic system and good points of applications for highly selective and hydrophilic mixed-mode have been established. further derivatizations. It comprised suitable applications for complex The incorporation of an, e.g., imidazole residue featured auspicious samples containing lipophilic aromatic or aliphatic frameworks as well characteristics including possible analyte interactions via hydrophobic as as opposing polar, acidic substance classes (Figure 1). well as hydrogen bonding or pH dependent electrostatic interactions. By the additional post-polymerization derivatization, imidazole was One aim of Huck´s lab is the development of non-invasive NIR portable Figure 1: HPLC chromatograms of a mustard seeds extract converted into a strong ion-exchanger which increasingly extended the measurement technology for the determination of optimum harvest before and after SPE using Poly(NVI/EGDMA). Highly acidic, water soluble glucosinolates are interacting with the application of solid-phase extraction in analytical platforms. time for medicinal plants (phytomics). With this approach it is for the quaternized imidazolium ring by ion-exchange interactions. In the field of vibrational spectroscopy the development of non-invasive, first time possible to detect within one measurement quality related Displacement during elution process is enabled by counter- ions of high relative binding strengths. fast techniques based on near-&mid-infrared (NIR&MIR), Raman chemical parameters such as the concentration of distinct ingredients spectroscopy plays a crucial role enabling simultaneous analysis of physical and also physical parameters, e.g. anti-oxidative and or anti-bacterial and chemical parameters, respectively. The development of miniaturized activity, respectively. Thereby, reference analytical methods as described portable spectrometers is highly useful. Imaging and mapping MIR/NIR/ in the prior paragraph play a crucial role next to quantum chemical Raman methods are developed for screening of sample composition with >> Research Grants spectrum simulation and 2D-COS (Figure 2). FFG GZ 859502; OEAD GZ ZA 07/2017; EPU 53/2016, a spatial resolution down to 1 µm, which enables the development of EPU 54/2016; Interreg GZ AB116; Interreg GZ ITAT1005; early diagnosis tools for cancer detection. Quantum chemical calculation BMWFW 402.000/003-II/6b/2012 Since the milk and horsemeat scandal it became clear that early food for spectrum simulation can provide substantial support especially in the >> Coworkers fraud detection is essential. Therefore, it is the aim to develop easy PD Dr. Rania Bakry, Assoz.-Prof. Dr. Matthias Rainer, interpretation of complex spectra. In case of low concentrated analytes applicable vibrational spectroscopic tools in order to determine the Prof.Dr. Sevgi Türker (visiting professor); Mag.Dr. the combination of selective enrichment techniques with IR and/or Cornelia Pezzei, Dr. Justina Grabska, Dr. Krzysztof Bec, origin and/or species of food such as apples, milk, cheese, meat and (postdoc); Mag. Lukas Bittner, Mag. Raphael Henn, Raman spectroscopy has been shown to be suitable. These techniques cereals. Additionally, NIR/MIR and especially Raman Imaging is highly Mag. Markus Huber, Stefanie Delueg MSc, Mag. Karl are termed as SEIRS (Surface enhanced infrared spectroscopy); MEIRS Handle, Mag. Christian Kirchler, Verena Wiedemair suitable for the discrimination of cancerous tissue from healthy with MSc., Anel Beganovic MSc., Sophia Mayr MSc. (Ph.D. (Material enhanced infrared spectroscopy) and SERS (Surface enhanced a resolution down to 1 µm. These techniques give promise that in due students); Andreas Agerer, Sebastian Lörcher (diploma Raman spectroscopy). students) time non-invasive light fiber based measurement can be applied. 22  Research summary Research summary  23

Characterization of RNA, proteins, and their noncovalent complexes by mass spectrometry

Kathrin Breuker

>> Department of Organic Chemistry

activated dissociation (CAD), thus allowing its use for probing tat binding sites in TAR RNA by top-down MS. At the same time, the MS data revealed time-dependent 1:2 and 1:1 stoichiometries of TAR-tat complexes and suggest structural rearrangements of TAR RNA induced by tat peptide binding.

Many RNA-protein and RNA-drug complexes involve interactions between guanidine and phosphate moieties. We have investigated >> Goal: A major focus of our research is to explore the determinants noncovalently bound complexes of an 8 nt RNA and six different of biomolecular structure, stability, binding, and dissociation in the gas ligands, all of which comprise a guanidinium moiety, by electrospray phase. Based on insight from fundamental and mechanistic studies, ionization (ESI) and CAD MS, and found that the order of complex we develop new methodology for protein and ribonucleic acid (RNA) stability correlated almost linearly with the number of ligand atoms characterization by mass spectrometry (MS), including the identification, Figure 3: Top-down MS using CAD can identify RNA that can potentially be involved in hydrogen bond or salt bridge modifications, localize modification sites, and reveal the localization, and relative quantitation of posttranslational and site-specific, relative extent of modification for each site in interactions with the RNA but not with the proton affinity (PA) of mixtures of RNA isomers or forms (H. Glasner, C. Riml, R. Figure 1: Native top-down MS reveals the sites of tat peptide posttranscriptional modifications, and the determination of the ligands. However, ligand dissociation in CAD of the complex ions Micura, K. Breuker Nucleic Acids Res. 45, 8014-8025, 2017) binding to TAR RNA in the 1:2 complex (E.M. Schneeberger, stoichiometry and binding sites of noncovalent complexes of RNA. was generally accompanied by proton transfer (PT) from protonated K. Breuker Angew. Chem. Int. Ed. 56, 1254-1258, 2017). ligand to deprotonated RNA, indicating conversion of salt bridge into >> Background: Mass spectrometry is an evolving technique with unique hydrogen bond interactions. The relative stabilities and dissociation potential for biomolecular characterization beyond mere sequencing. pathways of the (RNA+mL-nH)n- complexes of differing stoichiometry Current research in the field aims, for example, at developing “top- (m = 1-5) and net charge (n = 2-5) revealed both specific and unspecific down” and “middle-down” MS approaches for studying different ligand binding to the RNA. proteoforms, posttranscriptional modifications of non-coding RNA, and the higher order structure of functional biomolecular assemblies, for Nucleobase methylations are ubiquitous posttranscriptional which a solid understanding of biomolecular gas phase ion structure, modifications (PTMs) that can substantially increase the structural stability, binding, and dissociation is critical. diversity of RNA in a highly dynamic fashion with implications for gene expression and human disease. Especially for noncoding RNAs, >> Research Highlights and Outlook: RNA frequently associates with research progresses at a high rate but is still critically hindered by proteins in many biological processes. The characterization of RNA- lack of adequate methodology for the characterization of PTMs; high protein complex structures and binding interfaces by NMR spectroscopy, throughput deep sequencing does not generally provide information X-ray crystallography, or strategies based on chemical crosslinking, on PTMs. We have investigated how specific nucleobase methylations however, can be quite challenging. We have explored the use of an affect RNA ionization in ESI, and backbone cleavage in CAD and electron alternative method, native top-down mass spectrometry, for probing of detachment dissociation (EDD), and developed two new top-down MS

complex stoichiometry and protein binding sites at the single-residue >> Research Grants approaches for the characterization of RNA methylations in mixtures Figure 2: CAD MS of RNA-guanidine derivative complexes level of RNA. Our data showed that the electrostatic interactions FWF Y372, FWF P27347, FWF P30087 of either RNA isomers or nonisomeric RNA forms. Fragment ions were shows that their stability increases with the number of ligand atoms that can be involved in hydrogen bond or salt bridge between HIV-1 TAR RNA and a peptide comprising the arginine-rich >> Coworkers analyzed to identify the modification type, to localize the modification interactions, but not with the proton affinity of the ligands binding region of tat protein are sufficiently strong in the gas phase Heidelinde Glasner, Eva-Maria Schneeberger, Jovana sites, and to reveal the site-specific, relative extent of modification for (J. Vusurovic, E.M. Schneeberger, K. Breuker ChemistryOpen Vusurovic, Matthias Halper, Giovanni Calderisi, Simon 6, 739-750, 2017). to survive phosphodiester backbone cleavage of RNA by collisionally Chwatal each site. 24  Research summary Research summary  25

Gastropod metallothioneins in evolution: New rules for an old protein family

Reinhard Dallinger

>> Department of Zoology

>> Goal: To achieve an extensive perception of the potential and Figure 1: Flowchart of MT evolution in , showing by us for the first NMR structure of a three-domain MT from the marine the major gastropod subclasses (Patellogastropoda, significance of metallothionein (MT) genes and proteins for metal Vetigastropoda, Neritimorpha, and snail Littorina littorea in cooperation with the Oliver Zerbe group metabolism in animals by exploring their evolution and diversification Heterobranchia) on the left-hand site. Two-domain MTs of (University of Zürich) (Baumann et al. 2017). More recently, we discovered low or no metal specificity at all have so far been observed towards novel structures and functions in different animal lineages. in Patellogastropoda, Vetigastropoda and Neritimorpha a further three-domain Cd MT structure in elegans, a close (above, in blue). In contrast, metal-specific MT isoforms terrestrial relative of Littorian littorea (Schmielau et al., submitted). In (CdMTs and CuMTs) have evolved in Caenogastropoda and >> Background: For a long time, MT research has been focused on the role Heterobranchia (middle and lower part). Two-domain Cd- some other species of Caenogastropoda and Heterobranchia snails, the and significance of this protein family in vertebrates (mainly mammals) specific isoforms (as found inPomatias elegans) are ancestral Cd loading capacity has been boosted by invention of MT isoforms with and have given rise to higher-molecular MT structures by and a few selected model organisms. We believe, however, that the addition of a third (like in Littorina littorea) or even up to as much as 10 (ten!) Cd-binding domains, as recently discovered by us in true potential of a protein family can only be comprehended and ten (as in Alinda biplicata) Cd-binding domains (middle part, Alinda biplicata (Heterobranchia) (unpublished data, Figure 1). In most in red). Cu-specific MT isoforms have been derived in some acknowledged by encompassing its structural and functional diversity Heterobranchia by gene duplication from two-domain Cd- cases, Cd specificity at the protein level is accompanied by Cd-specific across all major organismic phyla. During the last years, a plethora of specific ancestors (lower part, in green). In some families of upregulation of respective CdMT genes. Interestingly, Cu-selective MT Heterobranchia, metal specificity has finally been lost again primary MT sequences has been provided from a variety of different by primary structure modification. isoforms have evolved from pre-existing Cd-specific ancestral variants bacteria, plants, fungi and animal lineages. Our group has contributed only in species of the Stylommatophora lineage (terrestrial snails in to a better understanding of structural and functional MT diversity by the clade of Heterobranchia) through gene duplication, followed by in-depth research of MT structures and functions in the mollusk clade structural modification towards Cu selectivity. This has been achieved, of Gastropoda (limpets, snails and slugs). These animals represent, in among others, by increasing in the primary structure of CuMTs the terms of evolutionary radiation, one of the most successful and diverse K : N (lysine : asparagine) ratio, which indicates that the selectivity animal phyla whose members have successfully adapted, several times towards distinct metal ions may at least in part be owing to structural independently, to life in terrestrial and freshwater habitats. One of the constraints affecting the ionic charge and size of metal binding pockets most challenging questions in this concern is, to which extent gastropod in the MT metal clusters (Palacios et al. 2011). A striking difference to MT structures and functions have been inherited from marine ancestors, other animal MTs is the evident lack of Zn2+ affinity in gastropod MTs. and by how much they may have undergone adaptive modifications Instead, in Gastropoda this metal ion is consistently associated with low- upon transition from sea to land and to freshwater environments. molecular metal binding ligands such as phytochelatins or carbohydrate compounds (unpublished data, in preparation). As an intriguing >> Research Highlights and Outlook: Through evolution of gastropod outlook, it becomes more and more apparent that one additional MTs, there has been a trend towards diversification into metal-specific and so far neglected functional significance of MTs in gastropods, and isoforms, with a particular prevalence, in some gastropod lineages (such perhaps in other animal clades too, may be their important protective as Stylommatophora snails), of Cd- and Cu-selective MTs (Palacios et al. >> Research Grants role in favor of Ca2+ pathways (unpublished data, in preparation). FWF I1482-N28 (DACH, leading agency: FWF); FWF 2011). Evidently, Cd specificity has evolved prior to Cu specificity and must I3032-B21 (DACH, leading agency: FWF) therefore, be regarded as the ancestral metal-specific feature, occurring References >> Coworkers today in species of both, Caenogastropoda and Heterobranchia snails Veronika Pedrini-Martha (postdoc); Reinhard Lackner Palacios et al. 2011, BMC Biology BMC 2011, 9:4, 1-20. (technical assistance); Michael Niederwanger, Martin (Figure 1). In some Cd-selective MTs, the capacity of metal loading has Baumann et al. 2017, Angew. Chem. Int. Ed. 56, 4617-4622. Dvorak, Raimund Schnegg (Ph.D. students); Lara Schmi- been boosted by addition of one Cd binding protein domain, as shown elau (diploma student) Schmielau et al. 2018, Sci. Tot. Environ., submitted. 26  Research summary Research summary  27

Radiation damage in biological compounds induced by low energy electrons

Stephan Denifl

>> Department of Ion Physics and Applied Physics

>> Goal: Exploring negative and positive ion formation by secondary electrons formed upon radiation of biological compounds.

>> Background: A large number of secondary particles are generated when energetic primary radiation (e.g. photons, ions or cosmic Figure 1: Electron ionization mass spectrum of isolated radiation) interacts with biological material like living cells. The most tetrahydrofuran (orange line) and tetrahydrofuran clusters (black line), respectively. Both spectra were recorded at the abundant secondary species formed are electrons which are released electron energy of about 70 eV. In case of clusters, a peak is with an average kinetic energy of a few eV. These electrons subsequently present at m/z 55, which can be assigned to the loss of OH from tetrahydrofuran. interact with cell components before they become a chemically inactive species. The electron interaction may however be severe even leading to single and double strand breaks of DNA. Therefore it is crucial to investigate the interaction of low energy electrons with simple biomolecules representing building blocks of biological material remains bound to an intact THF molecule due to the high proton affinity (nucleobases, amino acids, etc.). Mass spectrometry of anions formed of the molecule. Rather unexpected is the exclusive formation of (THF by electron attachment represents our experimental approach. – OH)+ from clusters, since in this case intramolecular bond cleavage is favored over intermolecular bond cleavage although substantial >> Research Highlights and Outlook: In recent experiments we carried out energy is deposited by the colliding electron. However, additional mass spectrometric studies with clusters of tetrahydrofuran. Studies in experiments with the deuterated molecule allowed an unambiguous the field of chemical physics often use tetrahydrofuran (THF) as model identification of this reaction channel, since we observed the loss of compound for the sugar moiety of DNA. The clusters were created by OD in this case. Studies with the deuterated compound also allowed the so-called supersonic expansion technique and crossed with a beam the identification of reaction pathways in the case of nanohydration. of electrons. Figure 1 shows the resulting mass spectrum below the In case of nanohydrated THF, fragmentation of the sugar is reduced in monomer, when THF clusters are ionized by electrons with the kinetic favor of fragmentation by the solvent molecules. energy of about 70 eV. Most of the ions formed correspond to those >> Research Grants when a single THF molecule is ionized. Only two exceptions can be Finally, we recently also intensified our studies with potential FWF-I1015, FWF-P22443 found: The protonated THF ion and the (THF – OH)+ ion are not found radiosensitizers developed for the improvement of radiation therapy. >> Coworkers in the mass spectrum for the single molecule. The former ion is easily Michael Neustetter (postdoc), Jusuf Khreis, Anita Ribar, For future studies on the molecular level, nitroimidazolic compounds Rebecca Meißner, Joao Ameixa (Ph.D. students), Katha- formed by fragmentation of a larger cluster, where the weak bond will be promising since those compounds have been successfully rina Fink, Bea Haslwanter, Julia Reitshammer (master between the THF moieties is broken upon the ionization, and a proton students) established for radiation therapy in Nordic countries. 28  Research summary Research summary  29

Regenerative potential of reprogrammed neural stem cells

Frank Edenhofer its maintenance in vitro. We found that Nanog enhances cell cycle progression of NIH 3T3, primary fibroblasts and ESCs by downregulation >> Department of Molecular Biology, Department of Genomics, Stem Cell Biology & of the cell cycle inhibitor p27KIP1 (also known as CDKN1B) establishing Regenerative Medicine a direct link between pluripotency establishment and cell cycle control.

The Myc family of transcription factors has been implicated in both >> Goal: To obtain human neural stem cells for disease modeling and i) the induction of artificial pluripotency in somatic cells, and ii) the cell therapy generation of a variety of human tumors. In a collaborative study with the group of Andreas Trumpp (German Cancer Research Center, >> Background: The Edenhofer group is working with mammalian Heidelberg) we functionally analyzed the role of Myc in pluripotent stem cells and has a long-standing interest in cellular regeneration stem cells. Deletion of both c-myc and N-myc strongly decreases and disease modeling particularly of neurological disorders. Cellular transcription, splicing, and protein synthesis, leading to a proliferation reprogramming enables the derivation of induced pluripotent stem Figure 1: Genetic ablation of Cx43/45 by Cre/loxP-mediated arrest. This study shows that Myc controls the biosynthetic machinery of mutagenesis disrupts primitive endoderm formation in cells (iPSCs) from somatic cells such as skin or blood cells. iPSCs are embryoid bodies (EBs). (A-F): Immunofluorescence analyses of stem cells without affecting their potency, thus regulating their entry functionally equivalent to embryonic stem cells (ESCs) i.e. they exhibit the primitive endoderm marker Gata6 and the pluripotency and exit from the dormant state. marker Nanog in EBs on day 6 of differentiation. Gata6 is an unlimited differentiation and self renewal potential. We aim at i) detectable in a substantial number of nuclei of 4-day-old understanding the transcriptional program of stemness properties, ii) Cx43/45 flox EBs (A-C) while it is only rarely found in Cx43/45- Cell fate specification of stem cells exiting the pluripotent state and deficient cells (D-F). (G-H): Control EBs on day 6 exhibit a exploring novel 2nd generation reprogramming paradigms and iii) well-structured surface layer of extraembryonic endoderm undergoing cell differentiation represents another key question that harnessing the potential of patient-specific reprogrammed cells for (Gata6) (G) and a-feto protein (AFP). Cells of the outer Gata6- our group addressed. In this respect we employed a 3D organoid model, and AFP-positive layer stained negative for the pluripotency biomedical applications such as disease modeling and cell therapy. marker Nanog. A defined outer layer of Gata6-positive cells embryoid bodies (EBs), to study the role of gap junctional intercellular was not observed in Cx43/45-deficient EBs (I). Scale: (A-F): 100 communication (GJIC) in early embryonic development. mm. Scale: (G-I): 50 mm. >> Research Highlights and Outlook: One aim of our lab is the elucidation Connexin (Cx) 43 and Cx45 are co-expressed in ESCs, form gap junctions of transcriptional regulation of self-renewal of mammalian stem cells. and are considered to exhibit adhesive function and/or to contribute Self-renewal belongs with differentiation to the major features of to the establishment of defined communication compartments. We stem cells that are closely associated with cell cycle progression. The generated Cx43/45-double deficient mouse ESCs and achieved almost molecular processes underlying the choice between cell division, >> Research Grants complete breakdown of GJIC. Cx43/45-dKO results in a block of FWF I 3029-B30, FWF W 1206-B18, bmwfw 10.420-WF/ differentiation and dormancy are uniquely regulated in stem cells and differentiation in EBs without affecting pluripotency and proliferation. V/3c/2016, DFG AOBJ 635622 represent a fundamental principle to control cell type specification, We further demonstrated that GJIC-incompetent ESCs fail to form >> Coworkers organ homeostasis, and potentially tumorigenesis. We explored the Katharina Günther, Katharina Kruszewski, Jerome primitive endoderm in EB cultures, representing the inductive key Mertens, Sandra Rizzi, Ahmad Salti (postdoc); Anita mechanistic role of transcriptional regulators Nanog and Myc in cell step of subsequent differentiation events. This work helps to gain Erharter, Gabriella Fenkart, Larissa Traxler (Ph.D. cycle control of pluripotent stem cells. Nanog is a homeodomain students); Veronika Fricke, Regina Gassler, Anna comprehensive understanding into mechanisms underlying early Hausruckinger, Felix Strasser (master students); Marta transcription factor that is necessary for the natural induction of lineage specification, which is essential in order to understand Suarez Cubero, Urban Tscheikner-Gratl (technicians); pluripotency in early mammalian development but dispensable for Caroline Baldemair, Marion Staudinger (secretary) embryonic development and stem cell based organoid cultures. 30  Research summary Research summary  31

Immuno-gerontology

Beatrix Grubeck-Loebenstein

>> Research Department for Biomedical Aging Research

>> Goal: To gain a better understanding of age-related changes within results demonstrate the role of inflammation and oxidative stress in age- the immune system in order to find new ways to prevent the loss of related changes of immune cell survival factors in the BM. Antioxidants immune function with age. may therefore be beneficial in counteracting immunosenescence by improving immunological memory in old age (Pangrazzi et al, Eur J >> Background: Age-related changes in T-lymphocytes, immune cells Immunol 2017;47:481-492). responsible for eliminating virus-infected cells and cancer cells from the body, are most detrimental. This is due to early degenerative changes Vaccination for the elderly: that take place in the thymus, the organ in which T cells mature. As the Vaccines against tetanus and diphtheria are among the most frequently thymus gradually loses its ability to replenish, the naïve T cells decrease used vaccines worldwide, but previous studies from our laboratory while memory and effector T cells increase in number and dominate have shown that protection against tetanus and particularly against the repertoire. This can lead to the loss of certain T cell specificities and diphtheria is unsatisfactory in adults and older persons. In the course changes in the composition of the T cell repertoire. One point of interest of the EU project “MARK-AGE” we analyzed tetanus- and diphtheria- in this respect is to explore the role of the bone marrow (BM) for the specific antibody concentrations in 2100 adults of different age from 6 maintenance of immunological memory in old age. Of practical relevance selected European countries (Austria, Belgium, Germany, Greece, Italy, is the question how vaccination can be optimized for the elderly. Poland) in order to investigate differences in the level of protection against tetanus and diphtheria across Europe. The data demonstrate >> Research Highlights and Outlook: that tetanus- and diphtheria-specific antibody concentrations vary The bone marrow (BM) and immunological memory in old age: greatly between countries, which is also reflected in the percentage The BM is important for the long-term maintenance of immunological of persons with antibody concentrations below the protective level memory, but the influence of aging on the production of survival (0.1IU/ml), which ranged from 2 to 31% for tetanus and 28-63% factors for effector/memory T cells and plasma cells in the human for diphtheria. In most countries, tetanus- and diphtheria-specific >> Research Grants BM has not yet been investigated. We could demonstrate that the antibody concentrations decrease with age. Tetanus-specific antibody EU HEALTH-F4-2011-280873, EU SSH-2012-1-320333, expression of molecules involved in the maintenance of immunological EU H2020-PHC-2014-2015-633964, EUREGIO-EFH, FWF concentrations are generally higher in males than in females, whereas ZFW012530, ÖAW-DOC 24089, TWF UNI-0404/2018, D. memory in the human BM changes with age. While IL-15 that protects no gender-related differences were found for diphtheria-specific Swarovski 2015/BIO-15, D. Swarovski 2016/BIO-22 + − antibodies. The studies demonstrate that the European population is potentially harmful CD8 CD28 T cells increases, IL-7 decreases. IL-6 is >> Coworkers also overexpressed. In contrast, APRIL, a plasma cell survival factor for B Carmen Giefing-Kröll, Stella Lukas Yani, Andreas Me- not satisfactorily protected against tetanus and diphtheria. Protection ryk, Birgit Weinberger (postdocs); Marco Grasse, Carina cells, is reduced. In contrast, IFN-y, TNF, and ROS accumulate in the BM should be improved by a life-long perspective on vaccination and more Miggitsch, Erin Naismith, Luca Pangrazzi (Ph.D. stu- in old age. IL-15 and IL-6 correlate with ROS levels in BM mononuclear dents); Daniel Breitenberger, Christina Putzer, Rebecca education to increase awareness of the necessity of regular vaccination Ralser (diploma students); Magdalena Hagen, Florian cells. IL-15 and IL-6 are also overexpressed in the BM of superoxide throughout life all over Europe (Weinberger B et al, Exp Gerontol 2017; Hatzmann, Franz Melzer (master students); Brigitte dismutase 1 knockout mice compared to their WT counterparts. Our Jenewein, Michael Keller (technicians) Oct 7. pii: S0531-5565(17)30516-8). 32  Research summary Research summary  33

Development of selectively acting antitumor drugs

Ronald Gust

>> Department of Pharmacy, Pharmaceutical Chemistry

>> Goal: Development of new antitumor drugs, which address new targets to overcome intrinsic and acquired resistance of tumor cells.

Figure 2: Intracellular localization in the cytoplasm of MCF-7 cells. >> Background: Nowadays cancer chemotherapy is yet accompanied by limiting unwanted side effects and the development of intrinsic or acquired resistance. Therefore, the search for new targets and novel COX content led to almost inactivity of the complexes against MCF-7 antitumor drugs are topics in the research of medicinal chemists. cells indicating that COX-2 is involved in the mode of action of the Especially, metal-based drugs as alternatives to cisplatin and carboplatin Co-ASS derivatives. are of high interest. They should address not the DNA but other targets in the tumor cells to circumvent cross-resistance. Of further interest Ironsalene complexes were designed to get a carrier-mediated uptake is the design of selective estrogen receptor modulators (SERM) with in tumor cells. Inside the cells, they generate an iron-dependent a unique binding mode at the estrogen receptor to get alternatives form of non-apoptotic cell death. This effect called ferroptosis is to tamoxifen with a safer hormonal profile. Therefore, we focussed morphologically, biochemically, and genetically distinct from apoptosis our interest on metal complexes, which i) inhibit the 2 and necrosis. It is induced by Fe(II/III) because of building of reactive (COX-2) over-expressed in mammary and colon carcinoma or ii) induce oxygen species and can be suppressed by the potent ferroptose- ferroptosis, an iron-dependent form of non-apoptotic cell death. Our inhibitor ferrostatin-1. With our compounds, it was demonstrated for new SERMs represent bivalent drugs with affinity to the ligand binding the first time that iron complexes can induce ferroptosis and free iron domain (LBD) and the coactivator-binding site to exclude agonistic ions are not a prerequisite. growth stimulating effects. Nuclear receptors play a crucial role in the development of hormone- >> Research Highlights and Outlook: In a preliminary structure activity dependent tumors. Tamoxifen and fulvestrant are promising drugs in study we could show that the acetyl salicylic acid derivative [prop-2- the hormone therapy of the ER-positive mammary carcinoma. They ynyl-2-acetoxybenzoate]dicobalthexacarbonyl (Co-ASS) represents prevent in different ways the binding of coregulators to the activated an unselective COX-1/2 inhibitor with cytotoxicity against breast estrogen receptors thereby causing antiestrogenic properties. Our cancer cells comparable to cisplatin. Exchange of the cobalt cluster approach targets both the ligand binding domain (LBD) and the by [ethylene]trichloroplatinate(II) (Zeise´s salt) strongly increased the coactivator-binding site. 4-Hydroxytamoxifen (4-OHT) or derivatives inhibitory effects at COX-1, but drastically reduced the COX-2 inhibition were linked by an alkyl spacer with small molecules, so called nuclear and the cytotoxicity. With the objective of increasing the selectivity for receptor alternate-site modulators (NRAMs), which competes with the Figure 1: Connection of the 4-hydroxytamoxifen derivative GW 7604 with an alkylpyrimidine NRAM . COX-2, we introduced a chlorine substituent in position 3, 4, 5, or 6 at coaktivator for its binding site (Figure 1). In this case, the LBD binder the ASS moiety of Co-ASS. This substituent utilizes on the one hand (e.g. 4-OHT) causes selectivity for the ER and the coactivator can the larger binding cave of COX-2 to prevent activity at this isoenzyme effectively be prevented from ER activation. Dependent on the used >> Research Grants and on the other hand it reduces the binding to COX-1 due to steric LBD binder, the dimers show high affinity to the ER only slightly lower FFG (West-Austrian BioNMR) repulsion. Indeed, all complexes retained their activity at COX-2 and than estradiol. High advantages are the fluorescence properties, which >> Coworkers were inactive at COX-1. In this context, we characterized the MDA-MB Benjamin Ma, Daniel Bäcker, Natalie Fahrner, Sina allow first investigations on the mode of action. The bivalent drugs are Götzfried, Alexandra Knox, Robert Mauersberger, 231 breast cancer as well as the HT-29 colon carcinoma cells as COX-1/2- vesicular accumulated in MCF-7 cells and were identified especially in Victoria Obermoser, Anna Schöpf, Alexander Weninger positive and the MCF-7 breast cancer cells as COX-negative. The missing (PhD students); Monika Cziferszky (postdoc) vesicles in the cytoplasm (Figure 2). 34  Research summary Research summary  35

Developmental biology and bioadhesion in basal animal model systems

Bert Hobmayer and Peter Ladurner

>> Department of Zoology

>> Goal: We focus on stem cell differentiation during embryonic of sea squirts. In addition, we now expand our search for bio-adhesives development, tissue turnover and regeneration, and analyze bio- to 20 different flatworm species. It is the goal to understand the adhesion in basal animal model organisms. mode of action of these molecules to enable the development of new synthetic counterparts. >> Background: By using simple model organisms such as cnidarian The cnidarian polyp Hydra and various flatworm species show an polyps, flatworms, and sea squirts, we study basic molecular and cellular unparalleled capacity for whole body regeneration. In order to understand processes of cell differentiation, pattern formation, and bioadhesion. cellular aspects of regeneration, the formation of organized tissue layers Of central interest are evolutionary conserved developmental pathways was analysed including changes in apical-basal and planar polarities of and master regulatory genes known to act throughout the animal epithelial cells at the closing wound. These behaviours are under molecular kingdom. Thereby, we aim at a better understanding of molecular control of conserved signalling pathways. We have concentrated on mechanisms and transfer our knowledge to higher organisms in order Figure 2: Live image of polarity changes in the actin the role of the Wnt/beta-Catenin signalling pathway. It is activated by cytoskeleton of Hydra epithelial cells in a bud just evaginating to point to potential targets for biomedical research. Furthermore, towards the right. initial wound healing and then orchestrates a regeneration-specific gene we have started to characterize bio-adhesive substances used by our regulatory network involved in position-specific differentiation events. animals to attach to the substrate. Regeneration is commonly based on the action of naïve, stem cell-like cells, and their behaviour is regulated by stemness-factors of the Myc Figure 1: Transgenic flatworms expressing fluorescent >> Research Grants proteins in different cell lineages. >> Research Highlights and Outlook: We have characterized adhesive FWF 25404; FWF 30347; J4071 Schrödinger-Program; family. The Hydra genome encodes for four myc gene family members Marie Curie FP7 626525; 4x ÖAW DOC Fellowships; proteins of the flatworm Macrostomum lignano using a transcriptomic similar to the genomes of vertebrates and mammals. In a long-going EU-COST Action Networks TD0906, CA15216, CA16203 and high throughput in situ screen approach combined with Mass effort and in strong collaboration with the CMBI group of Bister/Hartl/ >> Coworkers spectrometry, confocal- and electron microscopy, RNA interference, Ute Rothbächer (Associate Professor); Bernhard Egger Stefan, we study these ancestral forms of Myc factors, their structural (Assistant Professor); Marcelo Rodrigues (postdoc); specific antibodies and Lectin staining. Flatworms have evolved and biochemical properties in comparison to mammalian Myc factors, Birgit Lengerer, Robert Pjeta, Julia Wunderer, Willi adhesives adapted to different environments such as freshwater, Kari, Sabine Gufler, Fan Zheng (PhD students); Lena and their different roles in decision making in the interstitial stem cell Zitzelsberger, Belinda Artes, Veronika Prantl, Willi Sal- seawater and parasite attachment onto its host. We have started to lineage of Hydra. First data sets suggest actions in stem cell self-renewal, venmoser, Thomas Ostermann, Hermine Hohensinner identify bio-adhesives also in the cnidarian polyp Hydra and the larvae (technical assistants). cell multiplication, but also nerve cell differentiation. 36  Research summary Research summary  37

Molecular and cell biology of human aging

Pidder Jansen-Dürr In the Jansen-Dürr group, basic research and translational work is being >> Research Department for Biomedical Aging Research conducted on mechanisms of cellular senescence using model systems reflecting aging of the human skin To this end we study human cells >> Goal: The group studies molecular mechanisms of cellular senescence which are grown in vitro to either achieve replicative senescence through in human dermal fibroblasts and vascular endothelial cells, with a focus extended passaging (telomere shortening) or by using exogenous is on the role of reactive oxygen species,. In addition, we are interested Figure 2: UVB-induced changes in the morphology of recon- stressors to drive cells in stress-induced premature senescence (SIPS). We structed skin. Skin equivalents were submitted to UVB and in the role of mitochondrial proteins as regulators of senescence stained by hematoxylin–eosin for morphological analysis. have extended previous findings aiming at a better understanding of and in age-associated metabolic dysregulation. We also investigate molecular mechanisms underlying UVB-induced senescence of diploid the relationship between cellular senescence and age-associated human dermal fibroblasts (HDFs), an experimental model to study the dysfunctions and diseases, such as skin aging and cardiovascular process of photoaging of the skin. Our data suggest that autophagy diseases. Specific topics include the role of various stress signals on is required for the establishment of the senescent phenotype in UVB- cellular aging and age-associated pathologies of human tissues. We treated HDFs and that inhibition of autophagy is sufficient to change also developed a new diagnostic test system for detection of HPV E7 the cell fate from senescence to cell death by apoptosis (Fig. 1). proteins in cervical smears which was shown to be an efficient tool for triage of HPV-positive women. Studies in reconstructed skin equivalents revealed that UVB irradiation triggers hallmarks of autophagy induction in the dermal layer. These >> Background: The group has elucidated the role of oxidative stress findings have potential implications for fundamental as well as on senescence phenotypes of human endothelial cells and dermal translational research into skin aging, in particular photoaging. (Fig. 2). fibroblasts. To study molecular mechanisms underlying photoaging of the skin, we have used a model for UVB induced cellular senescence of In a second project we have continued to characterize the function Figure 3: Depletion of oxaloacetate decarboxylase FAHD1 human skin fibroblasts in which we identified protein quality control inhibits cell proliferation (A), induces cellular senescence in of FAHD1, a newly discovered metabolic enzyme which is the first mechanisms, such as proteasome activity and autophagy as new players human endothelial cells (B-C), and inhibits mitochondrial eukaryotic member of the family of oxaloacetate decarboxylases. The function (D-E). SA-ß-gal activity was increased in FAHD1 KD in cellular senescence of human dermal fibroblasts. We also developed cells (B). Western blots showing the expression of FAHD1, position of FAHD1 at the crossroads between carbohydrate metabolism a quasi-physiological model for photoaging of the skin, consisting p53, p21, and γ-H2AX (C). Mitochondrial membrane poten- and fatty acid metabolism suggests that FAHD1 plays an important role tial (MMP) (D) and oxygen consumption rate (OCR) (E) were of reconstructed human skin equivalents and used it to confirm the reduced in FAHD1 KD cells. as metabolic key regulator. Phenotypic alterations of FAHD1 KO mice role of protein quality control systems in skin aging. In addition, relative to wild-type mice are currently being investigated. In an attempt the mitochondrial protein FAHD1 was identified as a new player in to speed up this analysis we also knocked down FAHD1 in human regulation of mitochondrial function and cellular senescence; studies umbilical vein endothelial cells (HUVEC). We found that depletion of of molecular mechanisms are underway. FAHD1 from human endothelial cells inhibited mitochondrial energy metabolism and subsequently induced premature senescence. Whereas >> Research Highlights and Outlook: Human Aging is modulated by a senescence induced by FAHD1 depletion was not associated with >> Research Grants complex network of interacting genetic pathways, which have been DNA damage, we noted a reduction of mitochondrial ATP-coupled H2020 Marie Curie Actions RISE 691158 MediHealth, elucidated through studies on lower eukaryotic model organisms, EU FP7 305483 FRAILOMICS, Tiroler Wissenschaftsfond respiration associated with upregulation of the cdk inhibitor p21 (Fig. 3). UNI-0404/1625 ExomiR in OsteoAge, FFG Bridge 1 Figure 1: UVB induces autophagy. (A) LC3-GFP expressing such as yeast, flies and worms. However, not all mechanisms of aging These results provide additional support to the growing evidence that 848474 Alpine Pflanzen in der Kosmetik, Swarowski HDFs were submitted to UVB and processed for confocal identified in animal models can be faithfully and completely translated grants, industry grant Mikrogen GmbH, Neuried, Ger- mitochondrial dysfunction can induce cellular senescence by metabolic microscopy. Wild type HDFs were subjected to the same many, industry grant Cura Marketing GmbH, Innsbruck, treatment and analyzed by (B) Western blot or (C) electron into human aging. Accordingly, there is a need to study aging processes alterations independent of the DNA damage response pathway. Austria, AWS Grant FAHD1 Inhibitors, COST Action microscopy. (A) Representative images of three indepen- also in systems more closely mimicking human aging. In this respect, MITOEAGLE, PhD position LFUI Nachwuchsförderung In collaboration with Mikrogen GmbH in Neuried, Germany, and a dent experiments showing the punctuation of LC3 upon 1, 2, 3, and 4 days of UVB treatment (D1, D2, D3 and D4, the concept of cellular senescence has gained a lot of impact in the last >> Coworkers clinical partner from Theassaloniki, Greece, we evaluated high-risk(hr) respectively) and 3 days after the last UVB stress (D7). (B) Huaije Bu, Solmaz Etemad, Rafal Koziel, Maria Amalia decade primarily by the discoveries that i) cellular senescence occurs in HPVE7-protein detection as a triage method to colposcopy for hrHPV- Representative Western blot showing the lipidation of LC3 Cavinato Nascimento, Alexander Weiss (postdocs); Eva upon UVB, where LC3-I corresponds to unlipidated and LC3- aged organisms from primitive vertebrates up to humans and ii) the Albertini, Giorgia Baraldo, Michele Petit, Christina positive women, using a newly developed sandwich-ELISA-assay. This II to lipidated form of the protein. (C) Electron micrograph Metzger, Sophia Wedel (Ph.D. students); Max Holz- recent demonstration that the accumulation of senescent cells in mouse assay could be a useful partner to HPV testing for cervical cancer of cells submitted for 2 days to UVB irradiation and their knecht, Anne Buchmann (diploma students); Annabella respective controls. models drives the aging process. Pittl, Beata Szalka (technicians) screening. 38  Research summary Research summary  39

Plant biochemistry and metabolism

Ilse Kranner

>> Department of Botany

Figure 1: Schematic representation of the involvement of hydrogen peroxide (H2O2) and low-molecular- weight (LMW) thiols and disulphides during bread wheat germination and early seedling growth.

From left to right, changes in LMW thiol- disulphides and H2O2 production rates in Triticum aestivum: in a whole dry seed, seed structures isolated from non-germinated seeds after 15h from the onset of seed water uptake, and seed structures isolated from germinated seeds after 15 and 48h. Whole dry seed, endosperm including aleurone (large oval), and embryo or seedling (small oval) are divided by vertical lines. These lines delimit areas proportional to the concentrations of total glutathione [i.e. GSH (glutathione) + GSSG (glutathione disulphide)], area left of line) and cyst(e)ine [i.e. Cys (cysteine)

+ CySS, (cysteine) area right of line], in the respective seed structure. The redox states (Ehcs in mV) of total glutathione and cyst(e)ine are indicated by the blue-to-red (reducing-to-oxidising) shadings of

each area, as shown by the bottom right scale. Yellow background shadings indicate the rates of H2O2 production (nmol g-1 DW s-1), as shown by the bottom left scale. The dashed vertical line separates seeds from seedlings. Taken from Gerna et al. Free Radical Research 51:6, 568-581.

>> Goal: To deepen our understanding of plant metabolism with the aim to identify key molecular switchboards that determine plant stress response.

>> Background: Plants are the basis of life on Earth. All food is directly or indirectly derived from plants, and plants also produce the oxygen >> Research Highlights and Outlook: In this reporting period, much we breathe. In other words: no plants, no food, no oxygen. The current effort was directed towards delivering the € 3 million FP7 project climate change is accompanied by stress factors such as drought “Impacts of Environmental Conditions on Seed Quality” (EcoSeed). and elevated temperature - with potentially deleterious effects on Coordinated by the University of Innsbruck, the consortium involved agriculture and biodiversity, compromising food security for an ever- 11 partners from five European countries. The EcoSeed project was increasing human population. Our research group is dedicated to dedicated to unravelling the effects on seed quality of the stresses making significant contributions to unravelling the molecular basis of predicted to occur more frequently due to climate change: elevated plant stress response. In the past two years, much of our work focussed temperature and drought. Using a full omics approach complemented on research into the impacts of climate change on seed production of >> Research Grants by various biochemical and biophysical techniques, four main species crop plants. In addition, we worked on wild plant species from extreme EU FP7 grant 311840, EU FP7 Marie Curie grant 328370 were investigated, the model plant Arabidopsis thaliana, and the environments and of micro-algae, to further increase our understanding >> Coworkers crops Brassica oleracea, Helianthus annuus and Hordeum vulgare. A Thomas Roach; Wolfgang Stöggl (Assistant Professors); of plant response to environmental stress factors. We use mainly complementing study of bread wheat showed that seed germination Erwann Arc, Beatriz Fernández-Marín, Chae Sun Na, Fa- hyphenated techniques (UHPLC-MS/MS, GC-MS/MS) in combination bio Candotto Carniel (postdocs); Davide Gerna, Gregor is intricately regulated by thiol-disulphide conversions involving the Pichler (PhD students); Julian Wimmer, Theresa Baur with spectrophotometric methods to identify check points in plant tripeptide antioxidant glutathione (g-glutamyl-cysteinyl-glycine) in a (MSc students); Birgit Stenzel, Bettina Lehr, Siegfried metabolism that trigger cell, tissue and plant fate – to die or to survive. Aigner (technicians) tissue specific manner. 40  Research summary Research summary  41

Chemistry, chemical and structural biology of the pigments of life

Bernhard Kräutler and Thomas Müller

>> Department of Organic Chemistry

>> Goal: To gain knowledge on the chemistry, on biological roles and bio-molecular interactions of the porphyrinoid ‘pigments of life’, and to apply this in biology and medicine.

>> Background: A large part of our research concerns the ‘pigments of Figure 1: Natural Vitamin B12-derivatives are intimately as- Figure 2: Phyllochromobilins, the colored catabolites of chlo- life’, which have crucial and diverse roles in cells, e.g. as cofactors in sociated with their cobalt-center (the element cobalt was rophyll, are components of the fall colors in the leaves of de- enzymes, in biological processes driven by solar light and as metabolites named by German miners as the undesirable contamination ciduous tree put into various ores by a mine ‘Kobold’). regulating expression of genes. They comprise the porphyrinoids (heme, chlorophyll, and corrinoids) and linear tetrapyrroles, which

result from breakdown of heme and of chlorophyll. Porphyrinoids owe crystal structure was the first of a metal analogue of a natural 12B -

their important roles in Nature to their unique molecular properties derivative. The ‘mother’ ligand of vitamin B12, hydrogenobyric acid, and some of their basic structures are assumed to have pre-biotic origin. gave first insights into the intriguing capacity of the ‘contracted’ corrin Frequently they are functional complements to biological macro- macrocycle for binding and activating transition metals. AdoRhbl and

molecules, as coenzymes or as structuring and regulating ligands, e.g., other ‘antivitamins B12’ are inhibitors of B12-dependent enzymes and

in riboswitches. Our approach is geared at providing chemical insights interfere with B12-riboswitches, as predicted.

for the natural biological roles of the ‘porphyrinoid pigments of life’, as Figure 3: 2D Localization and identification of a chlorophyll Our lab has also pioneered studies on the structure and chemistry of well as at exploring their effects in important cellular processes. degradation product in senescent leaves of fern using MAL- chlorophyll catabolites from higher plants, named ‘phyllobilins’. In the DI-TOF mass spectrometry imaging. context of such studies we have become interested in the colored natural >> Research Highlights and Outlook:Our lab has made key contributions ‘phyllochromobilins’, which are strikingly similar, structurally, to bilins derived from heme catabolism. Finding evidence for (still hypothetical) to the chemistry of vitamin B12-derivatives, in particular as part of physiological effects of phyllobilins in plants, animals and humans is our recent program on ‘antivitamins B12’, metabolically inert B12- derivatives. This research will help us learn more about mechanisms one of the goals of this research. Furthermore, we have developed >> Research Grants of B -dependent metabolic processes in microorganisms, animals and MALDI-TOF and DESI mass spectrometry imaging methodology for the 12 FWF P-28522, FWF P-28892, Sparkling Science project SPA/04-140/INDIAN SUMMER IN TYROL. 2D localization and identification of biologically relevant molecules in humans, and in looking at still enigmatic effects of B12-deficiency by B12- chemical biological approaches. In this context, we have developed a >> Coworkers biological tissue. The imaging studies on chlorophyll breakdown also Ch. Li, S. Moser, F. Widner (postdocs), S. Murtaza (ÖAD synthetic approach towards a group of B -mimics that contain transition involved students from local partner high schools (’Sparkling Science’ 12 guest scientist), T. Erhart, C. Kieninger C. Meisenbichler, M. Scherl, C. Vergeiner, St. Vergeiner (Ph.D. students), C. project). Exciting results as well as simple experiments in this context metals other than the B12’s own cobalt (Figure 1). A combination of Brenig, M. Huber, J. Mayr, P. Moser, S. Schatz, P. Singe- biological and chemical total synthesis was established. A first example were presented to a broader public at the “Lange Nacht der Forschung wald (diploma students), M. Schäfer (master student), G. Scherzer (chemo-technician). 2016”. was AdoRhbl, the unnatural rhodium analogue of coenzyme B12. Its 42  Research summary Research summary  43

Protein dynamics and biomolecular recognition

Klaus R. Liedl and Daniela Schuster

>> Department of General, Inorganic and Theoretical Chemistry >> Department of Pharmacy, Pharmaceutical Chemistry

independent research field. It not only enables a better understanding of experimental findings, but also creates insights in macromolecular properties inaccessible to experiments.

>> Research Highlights and Outlook: Hydrophobicity plays a key role in biomolecular recognition processes. Although there are various experimental and theoretical techniques which are widely used, they provide only little information about water dynamics and entropic effects. We developed a new metric which derives spatial hydrophobicity values directly from the dynamics of the water molecules in molecular dynamics simulations. Classical molecular dynamics simulations are currently restricted to the nanosecond to microsecond timescale because of hardware limitations. >> Goal: The focus of our work lies on the characterization of key Most of the available enhanced sampling techniques use a biased potential features of biomolecular recognition by employing a wide range of energy function, which makes it difficult to extract dynamical properties state of the art computational methods and developing new methods of the unbiased ensemble. Following up on our previously presented to extract and profile key traits of biomolecules. metric of the dihedral entropies we developed a metric which allows >> Research Grants us to obtain reweighted dihedral entropies from accelerated molecular FWF P26997, FWF P30565, FWF P30737, FWF P26782, >> Background: Molecular recognition is a vital aspect of nearly all FFG Bridge EARLYSNOW, FWF/TWF Lise-Meitner grant dynamics simulations. Thus, we can capture dynamics happening on the biological processes, however, understanding them on a microscopical Maren Podewitz, ÖAW DOC/Birgit Waldner, ÖAD millisecond timescale with microsecond long simulations. FR9/2017, ÖAD ICM-2017-07338 grant to Navista Sri level is not trivial. One of the most important findings of the last few Octa Ujiantari, CAPES Science Without Borders, bmwfw In close collaboration with the Tollinger group as well as the lab decades is the fact that biomolecules cannot be described as a static PRIZE prototype grant, Boehringer Ingelheim of Richard Weiss at the University of Salzburg we focus on the >> Coworkers entity. In other words, looking at a single static crystal structure is not characterization of the dynamical properties of plant pollen allergens. Maren Podewitz, Michael Schauperl, Veronika Temml enough to understand the biological mechanisms of a molecule. The (postdocs), Stefania Monteleone, Wang Yin, Birgit Employing our previously presented dihedral entropy metric, we were studying of the dynamics of the protein and describing it as a structural Waldner, Anna S. Kamenik, Johannes R. Löffler, Ursula able to link molecular flexibilities of the major birch pollen allergen Bet Kahler, Monica Fernandez Quintero, Johannes Kraml, ensemble is essential for understanding the proteins’ characteristics Florian Hofer, Muhammad Akram, Sonja Herdlinger, v 1a, captured with molecular dynamics simulations, to experimental and functions. Fabian Mayr, Navista Sri Octa Ujiantari (Ph.D. students), thermal and proteolytic stabilities. We currently investigate these Emanuel Ehmki, Melanie Wachter, Franz Waibl, Radu The use of theoretical methods such as molecular modelling, molecular Talmazan, Dennis Dinu, Theresa Steinacher, Elisabeth dynamics on extended timescales using our aforementioned metric of dynamics or docking has become more and more important in the past. Vorhofer, Erika Strieder, Ralph Kandel, Sandra Wagner, calculating reweighted dihedral entropies from accelerated molecular Christian Vieider, Alois Dalkner (MSc. Students), Alexan- It has long passed from being solely supplemental to experiments to an der Spinn, Lisa Retter (system administrator) dynamics simulations. 44  Research summary Research summary  45

Developmental biology

Dirk Meyer and Pia Aanstad

>> Department of Molecular Biology

>> Goal: To understand molecular mechanisms underlying regulation Pancreatic regeneration: We established new tools for efficient and coordination of fate specification, differentiation, proliferation conditional pancreatic cell ablation in zebrafish. Using these tools we and migration in vertebrate embryogenesis and regeneration, with a identified a novel precursor population for acinar cell regeneration and focus on gastrulation and pancreas formation. in collaboration with Stefan Kubicek (CeMM,/Vienna) we determined activities of the malaria drug artemether on improved beta-cell >> Background: Development of endodermal organs such as the pancreas regeneration. requires complex and still poorly understood interactions of signaling pathways in order to guarantee the temporary and spatially correct Islet development: High resolution imaging of emerging endocrine cells coordination of cell proliferation, migration and differentiation. In our revealed until now unappreciated dynamic motility. By applying novel research we study three specific aspects of endoderm formation by applying quantitative assays, we determined that this motility is regulated by genetics, molecular and in vivo imaging approaches in zebrafish: (1) the PI3K and GPCR signaling, and is important for islet formation. functions of TGF-beta regulated transcription factors in early endodermal induction, (2) the role and regulation of Hedgehog (Hh) signaling in Diabetes genes: Work on zebrafish homologs of the neonatal diabetes endodermal patterning, cell proliferation and cell migration and (3) the >> Research Grants and sacral agenesis gene MNX1 revealed irx1 as a direct Mnx target FWF P25659, FWF P27338, FWF P30038, TWF 236277, molecular mechanisms underlying pancreatic islet cell formation and and a requirement for Mnx/Irx interaction in kidney formation. Further DOC 24701 maturation during embryonic development and regeneration. investigations make use of our recently established pdx1 and mnx1 >> Coworkers Robin Kimmel (Assistant Professor); Patrick Fischer, zebrafish mutants as diabetes model to uncover mechanisms behind Armin Wilfinger (postdoc); Julia Freudenblum, Réka >> Research Highlights and Outlook: Hh-signaling: In collaboration long-term deleterious effects of hyperglycemia. In international Lorincz, Dominik Regele, Nicole Schmitner, Philipp with Eduard Stefan (Biochemistry) we showed that the orphan receptor Tschaikner, Onur Temocin (Ph.D. Students), Greta collaborative studies, we are defining the progression and mechanisms Ebnicher, Gerald Lerchbaumer, Fabian Martin (master Gpr161 localises the Hh-signalling inhibitor PKA to the primary cilium, of diabetes-induced effects on tissue damage and on regeneration students); Eve Holtorf, Dzenana Tufegdzic, Sonja Töch- the main site of Hh signal transduction. terle (technician) capacities. 46  Research summary Research summary  47

Synthesis, structure, and function of non-coding RNAs

Ronald Micura

>> Department of Organic Chemistry

constraints contributing to catalysis of these ribozymes by chemical, structural, and biochemical studies. One of the new self-cleaving RNAs contains the so-called pistol motif revealed by comparative genomic analysis; its precise biological function is yet unknown. Our recent crystal structure of a pre-catalytic state of this RNA shows guanosine G40 and adenosine A32 close to the G53–U54 cleavage site (see graphics a). While the N1 of G40 is within 3.4 Å of the G53 2’-OH group that attacks the scissile phosphate (see graphics b), thus suggesting a direct role in general acid–base catalysis, the function of A32 was less clear. We found evidence from atom-specific mutagenesis that neither the N1 nor N3 base positions of A32 are involved in catalysis. By contrast, the ribose 2’-OH of A32 turned out to be crucial for the proper positioning of G40 through a H-bond network that involves G42 as a >> Goal: To obtain an integrated understanding of RNA modifi­cation bridging unit between A32 and G40. We also found that disruption of and RNA mediated regula­tion and catalysis. the inner-sphere coordination of the active-site Mg2+ cation to N7 of G33 makes the ribozyme drastically slower. A mechanistic scenario with >> Background: For many years it was believed that there were only A32 playing a major structural role and hydrated Mg2+ playing a major a small number of non-protein-coding RNAs (ncRNAs) and that catalytic role in cleavage, was therefore disclosed. they (tRNAs, rRNAs, spliceosomal RNAs) were involved primarily in assembling the predominant protein macromolecules. Even large RNA Another aim of our lab is to understand the functional roles of RNA classes, such as snoRNAs and microRNAs, remained undetected. In in the cell. Consequently, it is essential to elucidate the dynamics of recent years, it became apparent that ncRNAs are numerous and that their production, processing and decay. A recent method for assessing their cellular functions – on their own or in complex with proteins – are mRNA dynamics is metabolic labeling with 4-thiouridine (4sU), followed diverse and important. Our lab aims at a comprehensive molecular by thio-selective attachment of affinity tags. Detection of labeled understanding of cellular processes involving ncRNAs, in particular of transcripts by affinity purification and hybridization to microarrays or gene regulation by riboswitches but also of traditional ncRNAs such by deep sequencing then reveals RNA expression levels. Our own efforts as encountered during ribosomal translation. Our lab has a major focused on the development of a novel sequencing method (TUC-seq) focus on the chemical synthesis of RNA allowing the introduction of that eliminates affinity purification and allows for direct assessment site-specific modifications, naturally occurring and artificial ones. This of 4sU-labeled RNA. It employs an OsO4-mediated transformation to enables us to evaluate their structure and function by a great diversity convert 4sU into cytosine. We exemplified the utility of the new method of chemical and biophysical methods, with a focus on chemical and for verification of endogenous 4sU in tRNAs and for the detection of biochemical probing techniques, fluorescence spectroscopy (including pulse-labeled mRNA of seven selected genes in mammalian cells to single molecule imaging), NMR spectroscopy, and X-ray crystallography. >> Research Grants determine the relative abundance of the new transcripts. The results FWF (I1040, P27947), SNF (Early Postdoc.Mobility), FFG (West-Austrian BioNMR) prove TUC-seq as a straight-forward and highly versatile method for >> Research Highlights and Outlook: The discovery of four novel self- >> Coworkers studies of cellular RNA dynamics.

cleaving ribozyme classes in the years 2014 and 2015 has opened a unique Jennifer Gebetsberger, Thomas Amort (postdocs); opportunity to undertake structure-function studies to comparatively Marina Frener, Lukas Jud, Sandro Neuner, Christian Riml, Nikola Vušurović, Catherina Gasser, Elisabeth Finally, the continuous development of synthetic methods for efficient assess the architectural diversity, catalytic cores and mechanism of Fuchs, Elisabeth Mairhofer, Christoph Falschlunger, Eva access to chemically modified RNA with novel functional properties is in action. To this end, we have provided insights into the topological Neuner, Josef Leiter, Maximilian Himmelstoß (Ph.D.); Daniel Fellner (technician) the center of our research interests. 48  Research summary Research summary  49

Cell physiology and gene regulation

Bernd Pelster, Adolf Sandbichler, Thorsten Schwerte

>> Department of Zoology

Abb. 2: Cadmium exposure leads to phosphorylation of ATF and an increased expression of wMT- 2. The expression of wMT-2 is, however, suppressed when Cd-exposed earthworms are in addition injured.

by affecting protein trafficking. Characterization of the phenotype of this mutant showed that it could be a disease model for the analysis of the genetic background for muscular dystrophy and heart dysfunction.

>> Goal: Our research aims at the analysis of molecular and structural In our research focusing on cellular redox balance and it‘s role in mechanisms as well as genetic control pathways of physiological circadian timekeeping and hypoxia stress response we established the phenomena. use of new and improved fluorescent protein sensors enabling us to measure the redox balance in different cellular organelles and their >> Background: The adaptation of animals to changing environmental contribution to whole cell redox equilibrium. The cell physiological conditions includes adaptations at the cellular and molecular level in consequences of redox alterations and their effect on cellular oxygen order to maintain homeostasis of energy metabolism, ion regulation consumption and glycolytic metabolism are being addressed. and acid base balance. Changing oxygen partial pressures, variable light regimes or exposure to toxicants, for example, are readily perceived by Our studies demonstrated that remote levels of heavy metal stress animals of different developmental stage. In our work we focus on the already cause epigenetic changes in earthworms, which persist even large scale changes in the overall gene expression patterns induced by after the stressor has long disappeared. Moreover, we were able to these environmental perturbations, typically resulting in characteristic reveal that wound repair suppresses specific detoxification mechanisms, modifications in metabolic defense reactions, activity patterns, oxygen which might demonstrate a link between stress response and immunity. transport capacities and overall metabolic or ion regulatory activity. Of particular interest are the control mechanisms guiding these expression During the spawning migration the European eel is exposed to extreme changes at the transcriptional and translational level. The sophisticated hydrostatic pressures, and in a process named silvering eels prepare for interconnection and interaction between the different regulatory this migration. Silvering encompasses significant modifications at the Abb. 1: (Top) Anguillicola crassus, ~70 parasites of a single >> Research Grants swimbladder in a 90 mm petri dish; European eel with pathways is addressed using appropriate invertebrate and vertebrate transcriptional level in swimbladder tissue to adjust gas permeability, FWF P26363-B25; FWF I2984-B25; §27 Projekt DB-Nr: position of the swimbladder (red), (Middle) swimbladder; model animals. The data, obtained at the cellular, molecular and organ 215529; Nachwuchsförderung Projekt Nr.: 226115; metabolism and ion transport capacities to elevated hydrostatic scale bar represents 10 mm, (Bottom) Enzymes and Förderung des Vizerektorats für Forschung Nr.: 235916; metabolites (blue) involved in ROS detoxification in gas level, are discussed with respect to the possible adaptational benefit for pressure. Infection of the swimbladder with the nematode Anguillicola Lise Meitner (M2262-BBL) gland cells to avoid various oxidative damages (red); the whole organism. crassus significantly impairs these silvering induced changes in superoxide dismutase (SOD), glutathione peroxidase (GPx), >> Coworkers glutathione reductase (GR), reduced glutathione (GSH), Martina Höckner (Associate Professor); Birgit Fiechtner, swimbladder tissue, which may compromise the spawning migration. oxidized glutathione (GSSG), malondialdehyde (MDA). Bettina Peer (technicians); Maja Šrut, Gabriel >> Research Highlights and Outlook: The mutant line POPDC1S201F of By an improved sequencing the genome assembly for the European eel Schneebauer, Sigrid Zobl (postdocs); Victoria Drechsel the zebrafish is characterized by muscular dystrophy and arrhythmia (PhD student) could be significantly improved. 50  Research summary Research summary  51

Structure-functional activity relationship investigations on ligands interacting with opioid receptors

Helmut Schmidhammer and Mariana Spetea

>> Department of Pharmacy, Pharmaceutical Chemistry

Figure 1: The KOR is a key target for developing pharmacotherapies for neuropsychiatric disorders. (A) Dysfunction of the KOR/dynorphin system is responsible for the development of neuropsychiatric disorders and their comorbidity. (B) Differential modulation (activation or inhibition) of the KOR is regarded as a promising strategy for developing therapies for pain, addiction, mood and neurological disorders. (C) Generation of a new class of KOR ligands as small molecules featuring a diphenethylamine scaffold, which evolved as G protein KOR biased agonists, partial agonists and antagonists.

between G protein-dependent (responsible for beneficial effects, e.g. analgesia) and β-arrestin2-dependent (responsible for adverse effects) pathways with important therapeutic implications.

>> Goal: To perform basic and applied research as theoretical and ex- >> Research Highlights and Outlook: Our research group has generated perimental work in order to advance the current understanding on new KOR ligands as small molecules featuring a diphenethylamine structure-functional activity relationships for ligands interacting with scaffold (Figure 1C). Multidisciplinary, synergistic approaches ranging opioid receptors. from molecular in silico and in vitro levels to in vivo systems were combined by linking bioinformatics and computational systems >> Background: The kappa opioid receptor (KOR) and its endogenous with biochemical, pharmacological and disease animal models. Our ligand, dynorphin, have received major attention in past years due to lab established for the new KOR ligands: (i) a KOR target-oriented their involvement in a variety of physiological and behavioral responses pharmacological profile (high affinity, selectivity, full/partial agonism (i.e. pain, stress, motivation, emotion, cognition, reward). Dysregulation or antagonism at the KOR, and biased signaling towards G protein of the KOR/dynorphin system is responsible for the development and activation), (ii) antinociceptive efficacy in thermal nociceptive and maintenance of neuropsychiatric conditions (Figure 1A). Preclinical visceral mouse pain models, (iii) antiseizure efficacy in prodynorphin- and clinical evidence indicates that this system contributes to symptom knockout mice, (iv) anticonvulsant efficacy in a mouse model of clusters that are shared by many neuropsychiatric disorders, and thus temporal lobe epilepsy, and (v) reduced propensity for unwanted to their comorbidity (Figure 1A). Differential modulation of the KOR behavioral and side effects (i.e. aversion, sedation, motor dysfunction). is regarded as a promising strategy for developing therapies for pain, Structural, molecular and functional mechanisms that can form the >> Research Grants drug addiction, mood disorders, neurological conditions, and itching basis for such an improvement of the benefit/risk index were analyzed FWF I2463, FWF P30430, TWF UNI-0404/1596 (TWF), skin diseases by either activating or blocking the receptor (Figure 1B). Aktion D. Swarovski KG 2014 P7400-029-011, ÖAD-WTZ in detail. The emerged findings underline the strong potential of the FR12/2016. Although KOR activation does not produce dependence, euphoria, or developed KOR ligands as new and improved therapeutics for human >> Coworkers leads to respiratory suppression, it induces dysphoria, sedation and diseases. Furthermore, the emerged G protein biased KOR ligands Aquilino Lantero, Stefan Noha (postdocs); Filippo psychotomimesis. The concept of biased signaling has come forward Erli, Maria Dumitrascuta (Ph.D. students); Michael may be exploited for use as research tools for understanding the basic Mairegger, Andreas Ritsch, Lea Schläfer, Birgit with the realization that the KOR activates G protein-dependent and biology of KOR signaling, and most important to have a tremendous Steinkellner, Dominik Bucher, Katja Walker, Lisa-Marie independent signaling networks. Biased ligands can alter the linkage Watzke (diploma students) prospective for innovative drug discovery strategies. 52  Research summary Research summary  53

Targeted proteolysis in human diseases and its impact on drug development and production a) After we had identified MID1 as an important player in the development of prostate cancer together with groups from the Medical Rainer Schneider and Bernhard Auer University of Innsbruck, Berlin, Bonn and Dundee, we were able to identify small substance modulators of MID1 which are potential new >> Department of Biochemistry drugs that could interfere with cancer and neurodegeneration.

We found a potential anti prostate cancer drug that ablates the translation enhancing effect of MID1 on AR-mRNA, namely Metformin, which is a safe drug that is used extensively since 60 years to treat type 2 diabetes mellitus. Our finding provides an explanation for the recently found inverse correlation between Metformin-treatment and cancer incidence. Figure 1: MID1 is a modulator of PP2Ac influencing several major players and pathways In further searches for small substances targeting the MID1 complex, we recently identified resveratrol, the famous polyphenol found in red wine, as an inhibitor of the MID1-pathway that can trigger the dephosphorylation of hyperphosphorylated tau as found in Alzheimer´s disease. This finding was achieved in cooperations with the groups of >> Goal: We are studying targeted proteolysis in a wide range of aspects Sybille Krauss at the DZNE in Bonn and Susann Schweiger in Mainz (1). from the human ubiquitin system to specific directed evolution of Together with an EU-consortium (ERANET NEURON) including groups proteases for the development and production of drugs to treat human from Milan, Berlin and Mainz we are studying several syndromes that diseases. are associated with altered MID1/mTOR signaling and present with epilepsy and/or intellectual disabilities. Together with the proteomics >> Background: One of our main research fields covers the structure platform from the Medical University of Innsbruck (Lindner group) and functions of the MID1 protein complex, an important player in the we try to elucidate altered expression profiles of hippocampal regulation of phosphorylation and translational control. The ubiquitin synaptosomes from different MID1- , TSC2- and Rett-syndrome mouse MID1 targets the catalytic subunit of Protein Phosphatase 2A models to decipher the impact of mTOR in these diseases. (PP2A) for proteolytic degradation. Lack of human MID1 causes Opitz syndrome (midline defect with cognitive impairment). Furthermore Furthermore, in cooperation with the department of Neurology at the MID1 has an impact on mTOR signaling, alterations of which are Medical University of Innsbruck we are engaged in several research involved in several intellectual disability and epilepsy syndromes. As projects concerning movement disorder diseases. In a Tyrolian case we are also involved in a study on a novel, potentially mTOR-related of myoclonus epilepsy we were now able to identify the respective epilepsy syndrome caused by a protease-mutation found in a family mutated gene, a protease which seems to influence mTOR signaling, in Tyrol, we are interested to study the impact mTOR exerts on the thus connecting another epilepsy syndrome with this pathway. local protein homeostasis in hippocampal synaptosomes of mice models reflecting such mTOR-related syndromes and how this process might be b) Under the framework of the Austrian Center of Industrial regulated by specific protease activities. Biotechnology (ACIB) we are working successfully on several projects Additionally, we are engaged in the development of biotechnological (strategic projects and company projects) in which we are developing >> Research Grants tools namely novel proteases to improve the biotechnological novel biotechnological tools for up- and downstream processing (2, FWF-TRP002330, FFG-ACIB(K2) P25.081 (Company production of biogenic drugs. Project), FFG-ACIB P25.111 (Strategic Project), EFACTS- patents pending). EU242193, FWF- I 1570-B13 (ERANET NEURON), MFF253

>> Research Highlights and Outlook: With our central research on the c) Additionally, we are using bioinformatic approaches to understand >> Coworkers structure and function of the MID1 complex and the study of movement Rainer Schneider (Project leader), Sandra Pfurtscheller, the structure/function/evolution relationships between proteins. In this Kamil Rolski, Petra Engele, Christina Kröß, Alexander disorder diseases as well as with our biotechnological/bioinformatic respect we are analyzing highly variable viral proteins (Coronaviruses) Mödlhammer, Kevin Vince (PhD students), Simon Hofer, approaches we achieved several major goals: Stefan Felderer, Bernhard Sprenger (diploma students) and peptides that modulate the immune system like Thymulin (3). 54  Research summary Research summary  55

Cell signaling in chronic CNS disorders

Nicolas Singewald, Alexandra Koschak and Jörg Striessnig

>> Department of Pharmacy, Pharmacology and Toxicology

Molecular Pharmacology group (J. Striessnig) The major research areas of the CMBI project leaders in the Department of >> Goal: This group tries to better understand the physiological and Pharmacology and Toxicology the pathophysiology of neuropsychiatric pathophysiological role of voltage gated Ca2+ channels (VGCCs) in disorders (in particular fear, anxiety, autism spectrum disorders and human disease. Recently, a major focus was on autism spectrum Parkinson’s Disease), retinal disorders and endocrine disease (such as disorders (ASD) and Parkinson’s Disease (PD). primary hyperaldosteronism and diabetes mellitus). Most of this work >> Background: Excessive Ca2+ signalling can be harmful for cell function is embedded in the FWF-funded Spezialforschungsbereich SFB F44 “Cell and survival. We have recently found that excessive cellular Ca2+ entry signaling in chronic CNS disorders” (2011-2019), the doctoral programs into neurons through voltage-gated Ca2+ channels may also cause CNS Molecular Cell Biology and Oncology (MCBO, 2005-2018), Signal Figure: The Figure shows Ca2+ inward current through diseases, such as migraine or autism. Cav1.3 L-type Ca2+ channels (stably expressed in HEK-293 Processing in Neurons (SPIN), the recently approved doc.funds network cells) during electrical activity simulating the continuous >> Research Highlights and Outlook: We have characterized sporadic “CavX” and a European Training Network. activity of a dopamine neuron in the substantia nigra of mutations in the CACNA1D gene in patients with autism with our the midbrain. We tested if isradipine, a Ca2+ channel blocker currently clinically used as antihypertensive, can inhibit without neurological disorders, such as epilepsy. From six such mutations Neuropharmacology group (N. Singewald) these channels at nanomolar concentrations (20 nM, 3 µM). in seven patients we now learned that all cause characteristic gating Inhibition of Cav1.3 channels occurred with IC values in the 50 2+ >> Goal: Our main aim is to identify novel drug targets that facilitate low nanomolar range that should allow the in vivo inhibition changes of the channel that allow enhanced Ca entry in neurons. anxiety- and fear inhibitory mechanisms, including fear extinction, to of these channels by very high therapeutic doses of this drug Although only few patients with these mutations are described, Ca2+ in patients. guide improved treatment strategies for different forms of anxiety channel blockers may be useful to prevent excessive signalling. We disorders, which are the most prevalent of all mental disorders. therefore tested if clinically used Ca2+ channel blockers inhibit these >> Background: Effective long-term treatment for fear and anxiety- (so-called Cav1.3 L-type) calcium channels during simulated neuronal related disorders is a continuing challenge. One emerging treatment activity (Figure). Our experiments suggest that the required therapeutic strategy is combining exposure-based cognitive behavioral therapy with dose would be slightly higher than for blood pressure lowering. This extinction-facilitating drugs. Current research identified facilitation of hypothesis is now tested in a new animal model containing one of dopaminergic signaling and/or modulation of epigenetic mechanism, these mutations. in particular histone deacetylases and microRNAs, as means to boost and normalize aberrant fear-inhibitory learning, which is common in Molecular sensory physiology (A. Koschak) human anxiety disorders. >> Goal: Mapping the role of L-type calcium channels in retinal diseases. >> Research Highlights and Outlook: On the basis of these results we >> Background: In photoreceptors Cav1.4 L-type calcium channels are now gaining detailed insight into which dopamine receptors and (LTCCs) serve as the predominant source for Ca2+ entry to allow sustained brain region(s) mediate the normalization of impaired fear extinction >> Research Grants release of glutamate at their synaptic sites. Mutations in the encoding FWF SFB-F4401, FWF SFB-F4402, FWF SFB-F4410, learning by using pharmacological, optogenetic and chemogenetic CACNA1F gene have been associated with a number of human retinal FWF P24785, FWF P25014, FWF P25375, FWF approaches. One of our prominent findings was that the microRNA W1101, FWF W1206, FWF P26881, FWF P27809, diseases, such as congenital stationary night blindness type 2 (CSNB2). FWF I2433, ITN Horizon 2020 “switchboard”, Tiroler mir-144 is involved in orchestrating the rescue of impaired extinction >> Research Highlights and Outlook: We investigate mice carrying a Wissenschaftsförderung (TWF) ZAP740036, Univ. of learning. We are currently establishing whether this and other Innsbruck (FLD #242170, #194449, #217264, #214976), CSNB2 mutation which reproduces the functional phenotype described County of Tyrol (FLD #225100, #152787), H. Lundbeck microRNAs can be used as bio-markers/novel therapeutic targets for in a family with the corresponding mutation. We showed that similar A/S, FWF P27852, FWF P28146, FWF I02215 the treatment of anxiety disorders. In addition, we aim to improve the to human CSNB2 both rod and cone signaling pathways are affected. >> Scientific coworkers (2016/2017) tolerability of exposure based therapy by novel non-sedative anxiolytic Petronel Tuluc, Thomas Fenzl, Alexandra Pinggera, We demonstrated a defect in neuronal wiring in diseased mice evident Nadja Hofer, Nadine J Ortner, Kathrin Kähler, Hartwig drugs which do not impair extinction learning. Neuropeptide S (NPS) as formation of aberrant synaptic contacts and show remodeling of Seitter, Karl Ebner, Maria Kharitonova, Simone Sartori, is one such promising candidate. We currently pursue identification Anupam Sah, Conor Murphy, Verena Maurer, Thomas second order neurons. We currently investigate if Cav1.4 might also Keil, Veronica Fontebasso, Sinead Rooney, Michael of NPS-receptor agonists by virtual screening and in-vitro studies in be functionally relevant also in bipolar cells by generating cell-type Oberhauser, Lucia Zanetti, Irem Kilicarslan, Anita Siller, collaboration with CMBI groups of H. Stuppner, D. Schuster and R. Gust. Eva Maria Fritz, Joseph Moreno Rius specific Cav1.4 knock out mice. 56  Research summary Research summary  57

quantitative determination of several phenolic acids in coffee beans. Pharmacognosy – combining traditional knowledge with innovation Two aspects render this study noteworthy: first, the unique chemical composition / structure of the monolith and second, the fact that CEC has barely been used for the analysis of small molecules. Markus Ganzera and Hermann Stuppner Another focus is the discovery of bioactive natural products by >> Department of Pharmacy, Pharmacognosy Figure 2: DC separation and structures of bioactive activity guided isolation. Recent examples are investigations on the compounds isolated from the roots of Doronicum proanthocyanidin fraction of Ephedra sinica, which is able to significantly austriacum. Marzocco, S., Adesso, S., Alilou, M., Stupppner, H., Schwaiger, S. Molecules 2017, 22/6, Nr. 1003, doi: 10.3390/ inhibit XIAP (X-linked inhibitor of apoptosis protein), or the identification molecules22061003. >> Goal: Based on the traditional use of medicinal plants from around of anti-inflammatory compounds (i.e. benzofuranes, fig. 2) in the roots of the globe, modern approaches concerning the identification and bioac- Doronicum austriacum. In particular, 6,12-dihydroxy-(−)-2S-tremetone (1) tivity of therein found natural products, their analysis in crude drugs, showed the highest activity in the performed experiments and reduced products or biological samples by innovative techniques, and the use of ROS (reactive oxygen species) production in macrophages at 10 μM by in-silico activity prediction tools are the main targets of research. approximately 50%. Comparable results were observed for nitric oxide- release and iNOS (inducible ) expression. >> Background: It is more likely that natural products show bioactivity, because their biosynthesis proceeds with sequential binding of the In order to permit a comprehensive insight in our metabolome and biosynthetic intermediates to different proteins. Thus, they are to monitor small differences caused by diseases, lifestyle or nutrition, promising leads for drug discovery, and not surprisingly the vast majority highly sophisticated, selective, and sensitive analytical techniques like of commercial drugs actually derive from them. Pharmacognosy, which (U)HPLC-QToF-MS or NMR spectroscopy in combination with powerful focuses on the study of medicinal drugs derived from plants, has multivariate statistical tools such as PCA or OPLS-DA are applied. Within evolved from its classical, microscopy based origin to a modern and an ongoing FFG funded K-project “VASCage – Gut, Diet, Microbiota interdisciplinary area of research within the last decades. Today the and Vascular Ageing” we work on the establishment and validation covered topics range from the isolation and structural characterization of improved sample preparation protocols for the body fluids of natural products, to the development of analytical tools for Figure 3: Results of NMR studies indicating that directly urine, serum and feces (fig. 3). They are evaluated using the above prepared human feces extracts (orange) provide more mentioned analytical and statistical techniques, and permit a conclusive their qualitative and quantitative assessment, studies investigating reproducible results than those obtained with multi step metabolism or pharmacological properties by conventional or procedures (blue). Moosmang, S., Pitscheider, M., Sturm, S., identification of diverse biomarkers. Seger, C., Tilg, H., Halabalaki, M., Stuppner H. Clin. Chim. Since 2016, we are also coordinating the project “MediHealth – Novel computer based approaches, or ecologically relevant questions. At Acta. In press, doi: 10.1016/j.cca.2017.10.029. the Department of Pharmacy / Pharmacognosy all of these aspects natural products for healthy ageing from Mediterranean diet and food are covered and numerous publications as well as many successfully plants of other global sources”, which is funded by the Commission conducted research projects indicate an outstanding position in natural of the European Community (Research and Innovation Staff Exchange products research. Experienced staff scientists, motivated PhD students (RISE), Marie Curie Actions). The project is based on a well-balanced Figure 1: Separation of six lactones in Piper methysticum and a broad range of available equipment (LC-MS, GC-MS, CE-MS, LC- exchange of researchers between 5 universities and 4 companies roots by SFC. Murauer, A., Ganzera, M. Planta Med. 2017, 83, 1053-1057, doi: 10.1055/s-0043-100632. NMR, SFC-MS, 3D-databases, etc.) are the reason for this situation. from European countries as well as 4 universities from non-European countries. The main goal of the MediHealth project (http://medihealth. >> Research Highlights and Outlook: One main target of research is the eu/) is to introduce a novel approach for the discovery of active >> Research Grants evaluation of alternative, innovative techniques for natural products FWF P296710, FWF P293050, FWF P26917, FWF P24168, agents of food plants from the Mediterranean diet and other global analysis. Supercritical Fluid Chromatography (SFC) showed to be an FWF T942, FFG Alpine Kosmezeutika P7400-012-048, FFG sources to promote healthy ageing. To achieve this goal, plants from VASCage P7400-012-047, BMWF P7400-012-052, EU-FP7 interesting option, as for example the first successful separation IAPP – NATPROTEC P7400-012-037, EU H2020-MSCA-RISE the Mediterranean diet and edible plants from Africa, Asia and South of coumarins in Angelica dahurica and Ammi visnaga, or that of six - MediHealth P7400-012-049, EUREGIO ZFIPN000550, America have been carefully selected and subjected to in silico, in vitro, EUREGIO ZFIPN000310 lactones in Piper methysticum (fig. 1) could be achieved. All methods in vivo and metabolism analysis. Bioactive plant constituents will be >> Coworkers impress with extremely fast separations in the range of a few minutes Stefan Schwaiger, Sonja Sturm, Birgit Waltenberger isolated and identified. Pharmacological profiling of bioactive natural only. In an FWF funded project, novel stationary phases for Capillary (research associates); Anja Hartmann, Bianka Siewert, products as well as identification and synthesis of their metabolites will Veronika Temml (postdocs); Nora Engels, Gibitz-Eisath Electrochromatography (CEC) are evaluated. In cooperation with the Nora, Gratl Verena, Stefanie Hofer, Stefan Loos, Fabian be carried out. Finally, process optimization studies will be performed Department for Analytical Chemistry and Radiochemistry, an innovative Mayr, Simon Moosmang, Adele Murauer, Benjamin to develop innovative nutraceuticals, dietary supplements or herbal Mutschlechner, Maria Orfanoudaki, Luca Pompermaier, zwitterionic monolithic phase was developed and applied for the Silvia Revoltella (Ph.D. students) medicinal products. 58  Research summary Research summary  59

Biomolecular NMR spectroscopy

Martin Tollinger and Christoph Kreutz

>> Department of Organic Chemistry

>> Goal: The experimental characterization of structure, dynamics and function of proteins and (ribo)nucleic acids.

>> Background: It has become increasingly clear in the last decade that

the three-dimensional structures of proteins and (ribo)nucleic acids Figure 2: Left: Three-dimensional solution NMR structure of the major apple allergen Mal d 1. This 17.5 kDa protein is responsible for allergic reactions to apples in are generally flexible. Typically, these biomolecules adapt a defined >70% of all birch pollen sensitized patients. Center: Fully assigned, 2-dimensional 1H15N NMR spectrum of Mal d 1. Right: Mal d 1 was first identified and isolated from Golden Delicious and Granny Smith apples. distribution of structures around the „ground state“ that is obtained by standard structure determination protocols. Structural flexibility is required for the biological function of a biomolecule per se. For example, recognition and binding of bioactive ligands, catalysis of enzymatic reactions, regulation of enzymatic turnover and processing of allosteric information all rely on structural flexibility. In our group we aim at measuring and understanding structural flexibility of proteins and (ribo)nucleic acids at atomic resolution using NMR spectroscopy. We determine solution structures by standard NMR techniques and complement these data by NMR relaxation measurements to provide a genuine description of how these molecules function. We specifically focus on the mechanisms of ligand recognition and binding, protein and RNA folding and stability, and enzymatic catalysis.

>> Research Highlights and Outlook: One focus at our laboratory is the characterization of functional dynamics of (ribo)nucleic acids. Another focus at our laboratory are allergenic proteins such as Bet In this context, we are advancing the current protocols for isotope v 1, which represents the main cause for allergic responses to birch (13C, 15N, 2H) labeling of DNA and RNA. This is especially important pollen. A substantial proportion of individuals who suffer from birch for the application of dynamic NMR experiments, such as saturation pollen allergy develop intolerance to different kinds of fruits, nuts and transfer and relaxation dispersion experiments, where simple spin vegetables due to immunologic cross-reactivity of Bet v 1-specific IgE topologies are a mandatory prerequisite. For this task, we combine antibodies with homologous proteins in these kinds of food. The most chemical synthesis and NMR in a unique way. Recently, a method to frequent cross-allergies are directed towards apples and hazelnuts. We >> Research Grants introduce isotope labeling patterns into larger RNAs comprising up FWF-P26550, FWF-P26849, FWF-P28725, FWF-P30370, have recently determined the three-dimensional structure of the major FWF-P31054, FFG-858017, ERDF-ITAT1013, OEAW DOC to 80 nucleotides was established in our group. We currently focus apple allergen Mal d 1 and are currently working on allergenic proteins fellowship, Alpine Research Center Obergurgl on probing the functional dynamics of catalytically active non-coding from hazelnut and peach. In future experiments, we are planning Figure 1: General work flow. Chemical synthesis is used >> Coworkers to introduce atom-specific stable isotope labels (e.g.13 C, RNAs, such as a group II intron construct or the pistol ribozyme. L. Ahammer, V. Dietrich, R. Eidelpes, S. Führer, S. to establish how naturally occurring isoforms of these proteins with highlighted in orange) into RNA precursors. After sequence Grutsch, M. Juen, J. Kremser, J. Ludescher, F. Nußbaumer, This will give unprecedented insights into the structural flexibility >95% sequence identity and identical three-dimensional ground state assembly, solution NMR experiments are carried out to probe R. Plangger, E. Strebitzer (Ph.D. students), K. Erharter, J. structure and function of the target RNA. underlying biological function. Unterhauser (master students) structures can elicit different immunologic responses. 60  Publications Publications  61

G. Bonn, C. Huck K. Bister, M. Hartl, E. Stefan Bec K, Grabska J, Ozaki Y, Huck CW. Influence of non-fundamental Stefan E, Troppmair J, Bister K. Targeting the architecture of deregu- modes on mid-infrared spectra anharmonic DFT study of aliphatic lated protein complexes in cancer. Adv. Protein. Chem. Struct. Biol. in ethers. J. Phys. Chem. A 121, 1412-1424 (2017). press (2018). (Published online 18 August 2017). Henn R, Kirchler CG, Grossgut ME, Huck CW. Comparison of sensitivity Gufler S, Artes B, Bielen H, Krainer I, Eder MK, Falschlunger J, Bollmann to artificial spectral errors and multivariate LOD in NIR spectroscopy – A, Ostermann T, Valovka T, Hartl M, Bister K, Technau U, Hobmayer B. Determining the performance of miniaturizations on melamine in milk b-Catenin acts in a position-independent regeneration response in the powder. Talanta 166, 109-118 (2017). simple eumetazoan Hydra. Dev. Biol. 433, 310-323 (2018). (Published Huck, CW. Selected latest applications of molecular spectroscopy in online 3 November 2017). natural product analysis. Phytochemistry Letters 20, 491-498 (2017). Hunter E, Bister K (eds). Viruses, Genes, and Cancer - Current Topics Kirchler CG, Pezzei CK, Bec KB, Henn R, Ishigaki M, Ozaki Y, Huck CW. in Microbiology and Immunology, Vol. 407. Springer International Critical Evaluation of NIR and ATR-IR Spectroscopic Quantifications of Publishing (2017). Rosmarinic Acid in Rosmarini folium Supported by Quantum Chemical Stefan E, Bister K. MYC and RAF: Key effectors in cellular signaling and Calculations. Planta Med. 83/12/13, 1076-1084 (2017). major drivers in human cancer. Curr. Top. Microbiol. Immunol. 407, 117- Murauer A, Bakry R, Schottenberger H, Huck CW, Ganzera, M. An in- 151 (2017). novative monolithic zwitterionic stationary phase for the separation of Raffeiner P, Schraffl A, Schwarz T, Röck R, Ledolter K, Hartl M, Konrat phenolic acids in coffee bean extracts by capillary electrochromatogra- R, Stefan E, Bister K. Calcium-dependent binding of Myc to calmodulin. phy. Anal. Chim. Acta 963, 136-142 (2017). Oncotarget 8, 3327-3343 (2017). Jabeen F, Najam-ul-Haq M, Rainer M, Huck CW, Bonn GK. In-Tip

publications | Torres-Quesada O, Mayrhofer JE, Stefan E. The many faces of compart- Lanthanum Oxide Monolith for the Enrichment of Phosphorylated mentalized PKA signalosomes. Cell Signal. 37, 1-11 (2017). Biomolecules. Anal. Chem. 89/19, 10232-10238 (2017). Torres-Quesada O, Röck R, Stefan E. Systematic quantification of cAMP- Pezzei CK, Schönbichler SA, Kirchler CG, Schmelzer J, Hussain S, Huck- controlled PKA interactions. Horm. Metab. Res. 49, 240-249 (2017). Pezzei VA, Popp M, Krolitzek J, Bonn GK, Huck CW. Application of Bruystens JG, Wu J, Fortezzo A, Del Rio J, Nielsen C, Blumenthal DK, benchtop and portable near-infrared spectrometers for predicting the Röck R, Stefan E, Taylor SS. Structure of a PKA RIα Recurrent Acrodys- optimum harvest time of Verbena officinalis. Talanta 169, 70-76 (2017). ostosis Mutant Explains Defective cAMP-Dependent Activation. J. Mol. Teshome DA, Rainer M, Noel JC, Schüßler G, Fuchs D, Bliem HR, Bonn Biol. 428, 4890-4904 (2016). GK. Chemical compositions of traditional alcoholic beverages and con- Bachmann VA, Mayrhofer JE, Ilouz R, Tschaikner PM, Raffeiner P, Röck sumers’ characteristics, Ethiopia. African Journal of Food Science 11/7, R, Courcelles M, Apelt F, Lu T, Baillie GS, Thibault P, Aanstad P, Stelzl U, 234-245 (2017). Taylor SS, Stefan E. Gpr161 anchoring of PKA consolidates GPCR and cAMP signaling. Proc. Natl. Acad. Sci. USA 113, 7786-7791 (2016). K. Breuker Rinaldi L, Delle Donne R, Sepe M, Porpora M, Garbi C, Chiuso F, Gallo Vušurović J, Schneeberger EM, Breuker K. Interactions of protonated A, Parisi S, Russo L, Bachmann V, Huber RG, Stefan E, Russo T, Feliciello guanidine and guanidine derivatives with multiply deprotonated RNA A. praja2 regulates KSR1 stability and mitogenic signaling. Cell Death probed by electrospray ionization and collisionally activated dissocia- Dis. 7: e2230 (2016). tion. ChemistryOpen 6, 739-750 (2017). Ijaz M, Bonengel S, Zupančič O, Yaqoob M, Hartl M, Hussain S, Huck CW, Kremser J, Strebitzer E, Plangger R, Juen MA, Nußbaumer F, Glasner H, Bernkop-Schnürch A. Development of oral self nano-emulsifying delivery Breuker K, Kreutz C. Chemical synthesis and NMR spectroscopy of long system(s) of lanreotide with improved stability against presystemic thiol- stable isotope labelled RNA. Chem. Commun. (Camb) 53, 12938-12941 disulfide exchange reactions. Expert Opin. Drug Deliv. 13, 923-929 (2016). (2017). Hartl M. The quest for targets executing MYC-dependent cell transfor- Skinner OS, McAnally MO, Van Duyne RP, Schatz GC, Breuker K, Comp- mation. Front. Oncol. 6, 132 (2016). ton PD, Kelleher NL. Native electron capture dissociation maps to Iron- Patents binding channels in horse spleen Ferritin. Anal. Chem. 89, 10711-10716 Stefan E, Röck R, Raffeiner P, Bachmann V. Quantification of an interac- (2017). tion between a Ras protein and a Raf protein. European Patent Office, Glasner H, Riml C, Micura R, Breuker K. Label-free, direct localization EP16191530.1 (2016). and relative quantitation of the RNA nucleobase methylations m6A, Stefan E, Mayrhofer J. Full-length kinase conformation reporter. Euro- m5C, m3U, and m5U by top-down mass spectrometry. Nucleic Acids Res. pean Patent Office, EP16165865.3 (2016). 45, 8014-8025 (2017). 62  Publications Publications  63

Schneeberger EM, Breuker K. Native top-down mass spectrometry of TAR RNA in complex with a wild-Type tat peptide for binding site map- ping. Angew. Chem. 129, 1274-1278 (2017); Angew. Chem. Int. Ed. Engl. 56, 1254-1258 (2017). Palacios Ò, Jiménez-Marti E, Niederwanger M, Gil-Moreno S, Zerbe O, Schennach M, Schneeberger EM, Breuker K. Unfolding and folding of Atrian S, Dallinger R, Capdevila M. Analysis of metal-binding features the three-helix bundle protein KIX in the absence of solvent. J. Am. Soc. of the wild type and two domain-truncated mutant variants of Litto- Mass Spectrom. 27, 1079-1088 (2016). rina littorea Metallothionein reveals its Cd-specific character.Int. J. Mol. Cabedo Martinez AI, Weinhäupl K, Lee WK, Wolff NA, Storch B, Żerko Sci. 2017, 18, 1452; doi:10.3390/ijms18071452, pp. 1-16 (2017). S, Konrat R, Koźmiński W, Breuker K, Thévenod F, Coudevylle N. Bio- Schmielau L, Dvorak M, Niederwanger M, Dobieszewski N, Pedrini- chemical and structural characterization of the interaction between Martha V, Ladurner P, Rodríguez-Guerra Pedregal J, Maréchal JD, Dal- the Siderocalin NGAL/LCN2 and the N-terminal domain of its endocytic linger R. Differential response to Cadmium exposure by expression of receptor SLC22A17. J. Biol. Chem. 291, 2917-2930 (2016). a two and a three-domain metallothionein isoform in the land winkle Hoernes TP, Clementi N, Faserl K, Glasner H, Breuker K, Lindner H, : valuating the marine heritage of a . Sci. Hüttenhofer A, Erlacher M. Nucleotide modifications within bacterial Tot. Envion., submitted. messenger RNAs regulate their translation and are able to rewire the genetic code. Nucleic Acids Res. 44, 852-862 (2016). S. Denifl Flamm AG, Le Roux AL, Mateos B, Díaz-Lobo M, Storch B, Breuker K, Huber SE, Smialek MA, Tanzer K, Denifl S. Dissociative electron attach- Konrat R, Pons M, Coudevylle N. N-lauroylation during the expression ment to the radiosensitizing chemotherapeutic agent hydroxyurea. J.

publications | of recombinant N-myristoylated proteins. Implications and solutions. Chem. Phys. 144, 224309 (2016). Chembiochem. 17, 82-89 (2016). Itälä E, Tanzer K, Granroth S, Kooser K, Denifl S, Kukk E. Fragmentation patterns of 4(5)-nitroimidazole and 1-methyl-5-nitroimidazole - The R. Dallinger effect of the methylation. J. Mass Spectrom. 52, 770–776 (2017). Pedrini-Martha V, Schnegg R, Baurand PE, de Vaufleury A, Dallinger R. Neustetter M, Mahmoodi-Darian M, Denifl S. Study of electron ioniza- The physiological role and toxicological significance of the non-metal- tion and fragmentation of non-hydrated and hydrated tetrahydrofuran selective Cadmium/Copper-Metallothionein isoform differ between clusters. J. Am. Soc. Mass Spectrom. 28, 866–872 (2017). embryonic and adult helicid snails. Comp.Biochem.Physiol. C: Toxicol- Ribar A, Fink K, Li Z, Ptasinska S, Carmichael I, Feketeová L, Denifl S. ogy & Pharmacology 199, 38-47 (2017). Stripping off hydrogens in imidazole triggered by the attachment of a Baumann C, Beil A, Jurt S, Niederwanger M, Palacios Ò, Capdevila M, single electron. Phys. Chem. Chem. Phys. 19, 6406-6415 (2017). Atrian S, Dallinger R, Zerbe O. Structural Adaptation of a Protein to Ribar A, Fink K, Probst M, Huber SE, Feketeová L, Denifl S. Isomer Selec- Increased Metal Stress: NMR Structure of a Marine Snail Metallothio- tivity in Low-Energy Electron Attachment to Nitroimidazoles. Chemistry nein with an Additional Domain. Angew. Chem. Int. Ed. 56, 4617-4622 23, 12892-12899 (2017). (2017). Schürmann R, Tanzer K, Dabkowska I, Denifl S, Bald I. Stability of the Benito D, Niederwanger M, Izagirre U, Dallinger R, Soto M. Successive Parent Anion of the Potential Radiosensitizer 8-Bromoadenine Formed onset of molecular, cellular and tissue-specific responses in midgut gland by Low-Energy (<3 eV) Electron Attachment. J Phys Chem B: Condensed of Littorina littorea exposed to sub-lethal cadmium concentrations. Int. Matter, Materials, Surfaces, Interfaces & Biophysical 121, 5730–5734 J. Mol. Sci. 18, 1815; doi: 10.3390/ijms18081815, pp. 1-26 (2017). (2017). Niederwanger M, Dvorak M, Schnegg R, Pedrini-Martha V, Bacher K, Bidoli M, Dallinger R. Challenging the metallothionein (MT) gene of F. Edenhofer Biomphalaria glabrata: unexpected response patterns due to cadmium Edenhofer F, Dobeš J, Vobořil M, Brabec T, Dobešová M, Čepková A, exposure and temperature stress. Int. J. Mol. Sci. 18, 1747; doi: 10.3390/ Klein L, Rajewsky K, Filipp D. A novel conditional Aire allele enables ijms18081747, pp. 1-15 (2017). cell-specific ablation of the immune tolerance regulator Aire. Eur. J. Niederwanger M, Calatayud S, Zerbe O, Atrian S, Albalat R, Capdevila Immunol., Epub ahead of print (2017). M, Palacios Ò, Dallinger R. Biomphalaria glabrata metallothionein: Forero A, Rivero O, Wäldchen S, Ku HP, Kiser DP, Gärtner Y, Penning- lacking metal specificity of the protein and missing gene upregulation ton LS, Waider J, Gaspar P, Jansch C, Edenhofer F, Resink TJ, Blum R, suggest metal sequestration by exchangeiInstead of through selective Sauer M, Lesch KP. Cadherin-13 Deficiency Increases Dorsal Raphe 5-HT binding. Int. J. Mol. Sci. 2017, 18, 1457; doi: 10.3390/ijms18071457, pp. Neuron Density and Prefrontal Cortex Innervation in the Mouse Brain. 1-15 (2017). Front. Cell. Neurosci. 11, 307 (2017). 64  Publications Publications  65

Kwok CK, Ueda Y, Kadari A, Günther K, Heron A, Schnitzler AC, Rook Weinberger B, Keller M, Putzer C, Breitenberger D, Koller B, Fiegl S, M, Edenhofer F. Scalable stirred suspension culture for the generation Moreno-Villanueva M, Bernhardt J, Franceschi C, Voutetakis K, Gonos of billions of human induced pluripotent stem cells using single-use ES, Hurme M, Sikora E, Toussaint O, Debacq-Chainiaux F, Grune T, bioreactors. J. Tiss. Eng. Regen. Med., in press (2017). Breusing N, Bürkle A, Grubeck-Loebenstein B. Protection against Teta- Appelt-Menzel A, Cubukova A, Günther K, Edenhofer F, Piontek J, nus and Diphtheria in Europe: The impact of age, gender and country Krause G, Stüber T, Walles H, Neuhaus W, Metzger M. Establishment of origin based on data from the MARK-AGE Study. Exp. Gerontol., of a Human Blood-Brain Barrier Co-culture Model Mimicking the pii: S0531-5565(17)30516-8 (2017). Neurovascular Unit Using Induced Pluri- and Multipotent Stem Cells. Weinberger B, Keller M, Grubeck-Loebenstein B. Long-term mainte- Stem Cell Reports 8(4), 894-906 (2017). nance of diphtheria-specific antibodies after booster vaccination is Wörsdörfer P, Bosen F, Gebhardt M, Russ N, Zimmermann K, Komla hampered by latent infection with Cytomegalovirus. Immun. Ageing Kessie D, Sekaran T, Egert A, Ergün S, Schorle H, Pfeifer A, Edenhofer 14, 16 (2017). F, Willecke K. Abrogation of Gap Junctional Communication in ES Cells Pangrazzi L, Naismith E, Meryk A, Keller M, Jenewein B, Trieb K, Results in a Disruption of Primitive Endoderm Formation in Embryoid Grubeck-Loebenstein B. Increased IL-15 production and accumulation Bodies. Stem Cells 35(4), 859-871 (2017). of highly differentiated CD8+ effector/memory T cells in the bone mar- Wörsdörfer P, Mekala SR, Bauer J, Edenhofer F, Kuerten S, Ergün S. row of persons with Cytomegalovirus. Front. Immunol. 8, 715 (2017). The vascular adventitia: An endogenous, omnipresent source of stem Pangrazzi L, Meryk A, Naismith E, Koziel R, Lair J, Krismer M, Trieb K, cells in the body. Pharmacol. Ther. 171, 13-29 (2016). Grubeck-Loebenstein B. “Inflamm-aging” influences immune cell sur- Günther K, Menzel AA, Kwok CK, Walles H, Metzger M, Edenhofer F. Rapid vival factors in human bone marrow. Eur. J. Immunol. 47, 481-492 (2017).

publications | Monolayer Neural Induction of induced Pluripotent Stem Cells Yields Stably Weinberger B. Immunosenescence: The importance of considering age Proliferating Neural Stem Cells. J. Stem Cell Res. Ther. 6, 341(2016). in health and disease. Clin. Exp. Immunol. 187, 1-3 (2017). Safina D, Schlitt F, Romeo R, Pflanzner T, Pietrzik CU, Narayanaswami Weinberger B. Adult vaccination against tetanus and diphtheria: The V, Edenhofer F, Faissner A. Low-density lipoprotein receptor-related European perspective. Clin. Exp. Immunol. 187, 93-99 (2017). protein 1 is a novel modulator of radial glia stem cell proliferation, Stuetz W, Weber D, Dollé MET, Jansen E, Grubeck-Loebenstein B, Fiegl survival, and differentiation. Glia. 64(8), 1363-80 (2016). S, Toussaint O, Bernhardt J, Gonos ES, Franceschi C, Sikora E, Moreno- Schulze M, Hoja S, Winner B, Winkler J, Edenhofer F, Riemenschneider Villanueva M, Breusing N, Grune T, Bürkle A. Plasma carotenoids, MJ. Model testing of PluriTest® with next-generation sequencing data. tocopherols, and retinol in the age-stratified (35-74 years) general Stem Cells Dev. 25(7), 569-71 (2016). population: Cross-sectional study in six European countries. Nutrients Münst B, Thier M, Winnemöller D, Helfen M, Thummer RP, Edenhofer 8, 614 (2016). F. Nanog induces suppression of senescence via down-regulation of Zlamy M, Almanzar G, Parson W, Schmidt C, Leierer J, Weinberger B, p27KIP1 expression. J. Cell. Sci. 129(5), 912-20 (2016). Jeller V, Unsinn K, Eyrich M, Würzner R, Prelog M. Efforts of the hu- Scognamiglio R, Cabezas-Wallscheid N, Thier MC, Altamura S, Reyes man immune system to maintain the peripheral CD8+ T cell compart- A, Prendergast ÁM, Baumgärtner D, Carnevalli LS, Atzberger A, Haas ment after childhood thymectomy. Immunity & Ageing 13, 3 (2016). S, von Paleske L, Boroviak T, Wörsdörfer P, Essers MA, Kloz U, Eisen- Pinti M, Appay V, Campisi J, Frasca D, Fülöp T, Sauce D, Larbi A, man RN, Edenhofer F, Bertone P, Huber W, van der Hoeven F, Smith Weinberger B, Cossarizza A. Aging of the immune system: Focus on A, Trumpp A. Myc Depletion Induces a Pluripotent Dormant State inflammation and vaccination. Eur. J. Immunol. 46, 2286-2301 (2016). Mimicking Diapause. Cell 164(4), 668–680 (2016). Grasse M, Meryk A, Schirmer M, Grubeck-Loebenstein B, Weinberger B. Booster vaccination against tetanus and diphtheria: Insufficient B. Grubeck-Loebenstein protection against diphtheria in young and elderly adults. Immun. Weinberger B. Vaccines for the elderly: Current use and future chal- Ageing 13, 26 (2016). lenges. Immun. Ageing, in press (accepted Dec 1, 2017). Ciccarone F, Malavolta M, Calabrese R, Guastafierro T, Bacalini MG, Giacconi R, Costarelli L, Piacenza F, Basso A, Bürkle A, Moreno-Villanueva Reale A, Franceschi C, Capri M, Hervonen A, Hurme M, Grubeck- M, Grune T, Weber D, Stuetz W, Gonos ES, Schön C, Grubeck-Loebenstein Loebenstein B, Koller B, Bernhardt J, Schön C, Slagboom PE, Toussaint B, Sikora E, Toussaint O, Debacq-Chainiaux F, Franceschi C, Hervonen O, Sikora E, Gonos ES, Breusing N, Grune T, Jansen E, Dollé M, A, Slagboom E, Ciccarone F, Zampieri M, Caiafa P, Jansen E, Dollé MET, Moreno-Villanueva M, Sindlinger T, Bürkle A, Zampieri M, Caiafa P. Breusing N, Mocchegiani E, Malavolta M. Zinc-induced metallothionein Age-dependent expression of DNMT1 and DNMT3B in PBMCs from a in centenarian offspring from a large European population: The MARK- large European population enrolled in the MARK-AGE study. Aging AGE project. J. Gerontol. A Biol. Sci. Med. Sci., Epub ahead of print (2017). Cell 15, 755-765 (2016). 66  Publications Publications  67

Weinberger B, Joos C, Reed SG, Coler R, Grubeck-Loebenstein B. The Lengerer B, Wunderer J, Pjeta R, Carta G, Kao D, Aboobaker A, Beisel stimulatory effect of the TLR4-mediated adjuvant glucopyranosyl lipid C, Berezikov E, Salvenmoser W, Ladurner P. Organ specific gene expres- A is well preserved in old age. Biogerontology 17, 177-187 (2016). sion in the regenerating tail of Macrostomum lignano. Dev. Biol. 433, Grubeck-Loebenstein B, Pangrazzi L. Role of the bone marrow for 448-460 (2017). adaptive immunity in old age. In: Handbook of Immunosenescence, 2nd Demeuldre M, Hennebert E, Bonneel M, Lengerer B, Van Dyck S, Wat- edition, Fulop T, Franceschi C, Hirokawa K, Pawelec G (eds), Springer, tiez R, Ladurner P, Flammang P. Mechanical adaptability of sea cucum- in press ber Cuvierian tubules involves a mutable collagenous tissue. J. Exp. Biol. Weinberger B. Effects of ageing on the vaccination response. In: The 220, 2108-2119 (2017). Ageing Immune System and Health. Bueno V, Lord J, Jackson T (eds), Aufschnaiter R, Wedlich-Söldner R, Zhang X, Hobmayer B. Apical and Springer International Publishing Switzerland 2017, pp 69-86. ISBN 978- basal epitheliomuscular F-actin dynamics during Hydra bud evagina- 3-319-43365-3 tion. Biol. Open 6, 1137-1148 (2017). Dobner J, Ress C, Rufinatscha K, Salzmann K, Salvenmoser W, Folie R. Gust S, Wieser V, Moser P, Weiss G, Goebel G, Tilg H, Kaser S. Fat-enriched Kaserer T, Obermoser V, Weninger A, Gust R, Schuster D. Evaluation rather than high-fructose diets promote whitening of adipose tissue in of selected 3D virtual screening tools for the prospective identification a sex-dependent manner. J. Nutrit. Biochem. 49, 22-29 (2017). of peroxisome proliferator-activated receptor (PPAR)γ partial agonists. Egger B, Bachmann L, Fromm B. Atp8 is in the ground pattern of flat- European Journal of Medicinal Chemistry 124, 49-62 (2016). worm mitochondrial genomes. BMC Genomics 18, 414 (2017). Obermoser V, Urban ME, Murgueitio MS, Wolber G, Gust R. New Farkas J, Salaberria I, Styrishave B, Staňková R, Ciesielski TM, Olsen AJ,

publications | telmisartan-derived PPAR γ agonists: impact of the 3D-binding mode Posch W, Flaten TP, Krøkje Å, Salvenmoser W, Jenssen BM. Exposure on the pharmacological profile. European Journal of Medicinal of juvenile turbot (Scophthalmus maximus) to silver nanoparticles and Chemistry 124, 138-152 (2016). 17α-ethinylestradiol mixtures: Implications for contaminant uptake and Kinthada P, Gust R. Synthesis and characterization, nuclease activity plasma steroid hormone levels. Envir. Pollution A 220, 328-336 (2017). studies and anticancer activity of Au(III)isostatinthiosemicarbazone Gammoudi M, Garbouj M, Egger B, Tekaya S. Updated inventory and complexes as potential anticancer drugs. World Journal of Pharmacy distribution of free-living flatworms from Tunisian waters. Zootaxa and Pharmaceutical Science 8, 727-739 (2016). 4263, 120-138 (2017). Liu W, Gust R. Update on metal N-heterocyclic carbene complexes as Krasnokutski SA, Goulart M, Gordon EB, Ritsch A, Jäger C, Rastogi M, potential anti-tumor metallodrugs. Coordination Chemical Reviews Salvenmoser W, Henning T, Scheier P. Low-temperature condensation 329, 191-213 (2016). of carbon. Astrophys. J. 847, 89 (2017). Obermoser V, Mauersberger R, Schuster D, Czifersky M, Lipova M, Robertson HE, Lapraz F, Egger B, Telford MJ, Schiffer PH. The mito- Siegl M, Kintscher U, Gust R. Importance of 5/6-aryl substitution on chondrial genomes of the acoelomorph worms Paratomella rubra, the pharmacological profile of 4´-((2-propyl-1H-benzo[d]imidazol-1-yl) Isodiametra pulchra and Archaphanostoma ylvae. Scientific Reports 7, methyl)-[1,1´-biphenyl]-2-carboxylic acid derived PPARγ agonists. 1847 (2017). European Journal of Medicinal Chemistry 126, 590-603 (2017). Rodrigues M, Ostermann T, Kremeser L, Lindner H, Beisel C, Berezikov Maia P I da S, Carneiro Z A, Lopes C D, Oliveira C G, Silva J S, de Albu- E, Hobmayer B, Ladurner P. Profiling of adhesive-related genes in the querque S, Hagenbach A, Gust R, Deflon V M, Abram U. Organometal- freshwater cnidarian Hydra magnipapillata by transcriptomics and lic gold(III) complexes with hybrid SNS-donating thiosemicarbazone proteomics. Biofouling 32, 1115-1129 (2016). ligands: cytotoxicity and anti-trypanosoma cruzi activity. Dalton Lengerer B, Flammang P, Salvenmoser W, Ladurner P. Adhesive Organ Transaction 46, 2559-2571 (2017). regeneration in Macrostomum lignano. BMC Dev. Biol. 16, 20 (2016). Plusquin M, De Mulder K, Van Belleghem F, DeGheselle O, Pirotte N, B. Hobmayer, P. Ladurner Willems M, Cuypers A, Salvenmoser W, Ladurner P, Artois T, Smeets Wudarski J, Simanov D, de Mulder K, Ustyantsev K, Grelling M, Grud- K. Toxic effects of cadmium on flatworm stem cell dynamics: A tran- niewska M, Beltman F, Glazenburg L, Demircan T, Wunderer J, Qi W, scriptomic and ultrastructural elucidation of underlying mechanisms. Vizoso DB, Weissert PM, Olivieri D, Mouton S, Guryev V, Aboobaker A, Environ. Toxicol. 31, 1217-1228 (2016). Scharer L, Ladurner P, Berezikov E. Efficient transgenesis and annotated Rivera-Ingraham GA, Nommick A, Blondeau-Bidet E, Ladurner P, Lignot genome sequence of the regenerative flatworm model Macrostomum JH. Salinity stress from the perspective of the energy-redox axis: Lessons lignano. Nature Comm. 8, 2120 (2017). from a marine intertidal flatworm. Redox Biol. 10, 53-64 (2016). 68  Publications Publications  69

Agorastos T, Chatzistamatiou K, Moysiadis T, Kaufmann AM, Skenderi A, Lekka I, Koch I, Soutschek E, Boecher O, Kilintzis V, Angelidou S, Katsiki E, Hagemann I, Boschetti Gruetzmacher E, Tsertanidou A, Ange- lis L, Maglaveras N, Jansen-Duerr P. Human papillomavirus E7 protein Rodrigues M, Leclère P, Flammang P, Hess M W, Salvenmoser W, Hob- detection as a method of triage to colposcopy of HPV positive women, mayer B, Ladurner P. The cellular basis of bioadhesion of the freshwater in comparison to genotyping and cytology. Final results of the PIPAVIR polyp Hydra. BMC Zoology 1, 3 (2016). study. Int. J. Cancer 141(3), 519-530 (2017). Seybold A, Salvenmoser W, Hobmayer B. Sequential development of Chatzistamatiou K, Moysiadis T, Angelis E, Kaufmann A, Skenderi A, apical-basal and planar polarity in aggregating epitheliomuscular cells Jansen-Duerr P, Lekka I, Kilintzis V, Angelidou S, Katsiki E, Hagemann I, in Hydra. Dev. Biol. 412, 148-159 (2016). Tsertanidou A, Koch I, Boecher O, Soutschek E, Maglaveras N, Agoras- Gammoudi M, Salvenmoser W, Harrath AH, Tekaya S, Egger B. Ultra- tos T. Diagnostic accuracy of high-risk HPV DNA genotyping for primary structure of spermatogenesis and mature spermatozoa in the flatworm cervical cancer screening and triage of HPV-positive women, compared Prosthiostomum siphunculus (Polycladida, Cotylea). Cell Biol. Internat. to cytology: preliminary results of the PIPAVIR study. Arch. Gynecol. 40, 277-288 (2016). Obstet. 295(5),1247-1257 (2017). Gammoudi M, Salvenmoser W, Tekaya S, Egger B. Ultrastructure of Petit M, Koziel R, Etemad S, Pircher H, Jansen-Dürr P. Depletion of the ovary and oogenesis in the flatworm Prosthiostomum siphunculus oxaloacetate decarboxylase FAHD1 inhibits mitochondrial electron (Polycladida, Cotylea). Cell Biol. Internat. 40, 1174-1186 (2016). transport and induces cellular senescence in human endothelial cells. Kari W, Zeng F, Zitzelsberger L, Will JP, Rothbächer U. Embryo micro- Exp. Gerontol. 92, 7-12 (2017). injection and electroporation in the chordate Ciona intestinalis. JOVE Khan A, Dellago H, Terlecki-Zaniewicz L, Karbiener M, Weilner S, 116, e54313 (2016). Hildner F, Steininger V, Gabriel C, Mück C, Jansen-Dürr P, Hacobian A, Ott E, Wendik B, Srivastava M, Pacho F, Töchterle S, Salvenmoser W, Scheideler M, Grillari-Voglauer R, Schosserer M, Grillari J. SNEVhPrp19/ Meyer D. Pronephric tubule morphogenesis in zebrafish depends on hPso4 Regulates Adipogenesis of Human Adipose Stromal Cells. Stem

publications | Mnx mediated repression of irx1b within the intermediate mesoderm. Cell Reports 8(1), 21-29 (2017). Dev. Biol. 411, 101-114 (2016). Weilner S, Schraml E, Wieser M, Messner P, Schneider K, Wassermann Zobl S, Salvenmoser W, Schwerte T, Gebeshuber I, Schreiner M. Morpho K, Micutkova L, Fortschegger K, Maier AB, Westendorp R, Resch H, peleides butterfly wing imprints as structural colour stamp. Bioinsp. Wolbank S, Redl H, Jansen-Durr P, Pietschmann P, Grillari-Voglauer R, Biomim. 11, 016006 (2016). and Grillari J. Secreted microvesicular miR-31 inhibits osteogenic differ- Oda-Ishii I, Kubo A, Kari W, Suzuki N, Rothbächer U, Satou Y. A ma- entiation of mesenchymal stem cells. Aging Cell 15 (4), 744-54 (2016). ternal system initiating the zygotic developmental program through Weiher H, Pircher H, Jansen-Durr P, Hegenbarth S, Knolle P, Grunau S, combinatorial repression in the ascidian embryo. PLoS Genetics 12, Vapola M, Hiltunen JK, Zwacka RM, Schmelzer E, Reumann K, and Will e1006045 (2016). H. A monoclonal antibody raised against bacterially expressed MPV17 Egger B. Making heads or tails of tapeworms. Trends Parasit. 32, 511- sequences shows peroxisomal, endosomal and lysosomal localisation in 512 (2016). U2OS cells. BMC Res. Notes 9, 128 (2016). Juliano C, Hobmayer B. Animal evolution: New perspectives from early Prior KK, Wittig I, Leisegang MS, Groenendyk J, Weissmann N, Michalak emerging metazoans. BioEssays 38, 216-219 (2016). M, Jansen-Durr P, Shah AM, and Brandes RP. The Endoplasmic Reticu- lum Chaperone Calnexin Is a NADPH Oxidase NOX4 Interacting Protein. P. Jansen-Dürr J. Biol. Chem. 291(13), 7045-59 (2016). Meitzler JL, Makhlouf HR, Antony S, Wu Y, Butcher D, Jiang G, Juhasz A, Nlandu-Khodo S, Dissard R, Hasler U, Schafer M, Pircher H, Jansen-Durr Lu J, Dahan I, Jansen-Dürr P, Pircher H, Shah AM, Roy K, Doroshow JH. P, Krause KH, Martin PY, and de Seigneux S. NADPH oxidase 4 deficiency Decoding NADPH oxidase 4 expression in human tumors. Redox Biol. increases tubular cell death during acute ischemic reperfusion injury. 13, 182-195 (2017). Sci. Rep. 6, 38598 (2016). Kofler B, Borena W, Manzl C, Dudas J, Wegscheider AS, Jansen-Dürr P, Holl M, Koziel R, Schafer G, Pircher H, Pauck A, Hermann M, Klocker Schartinger V, Riechelmann H. Sensitivity of tumor surface brushings H, Jansen-Durr P, and Sampson N. ROS signaling by NADPH oxidase 5 to detect human papilloma virus DNA in head and neck cancer. Oral modulates the proliferation and survival of prostate carcinoma cells. Oncol. 67,103-108 (2017). Mol. Carcinog. 55 (1), 27-39 (2016). 70  Publications Publications  71

Cavinato M, Koziel R, Romani N, Weinmullner R, Jenewein B, Hermann M, Dubrac S, Ratzinger G, Grillari J, Schmuth M, and Jansen-Durr P. Ganthaler A, Stöggl W, Mayr S, Kranner I, Schüler S, Wischnitzki E, Sehr UVB-Induced Senescence of Human Dermal Fibroblasts Involves Impair- EM, Fluch S, Trujillo-Moya C. Association genetics of phenolic needle ment of Proteasome and Enhanced Autophagic Activity. J. Gerontol. A compounds in Norway spruce with variable susceptibility to needle Biol. Sci. Med. Sci. 72 (5), 632-639 (2016). bladder rust. Plant Mol. Biol. 94, 229-251 (2017). Bu H, Baraldo G, Lepperdinger G, and Jansen-Durr P. mir-24 activity Aigner S, Holzinger A, Karsten U, Kranner I. The freshwater red alga propagates stress-induced senescence by down regulating DNA topoi- Batrachospermum turfosum (Florideophyceae) can acclimate to a wide somerase 1. Exp. Gerontol. 75, 48-52 (2016). range of light and temperature conditions. Eur. J. Phycol. 52, 238-249 Cavinato M, Waltenberger B, Baraldo G, Grade CVC, Stuppner H, (2017). Jansen-Dürr P. Plant extracts and natural compounds used against UVB- Gerna D, Roach T, Stöggl W, Wagner J, Vaccino P, Limonta M, Kranner induced photoaging. Biogerontology 18(4), 499-516 (2017). I. Changes in low-molecular-weight thiol-disulphide redox couples are Bu H, Wedel S, Cavinato M, Jansen-Dürr P. MicroRNA Regulation of part of bread wheat seed germination and early seedling growth. Free Oxidative Stress-Induced Cellular Senescence. Oxid. Med. Cell. Longev. Radical Res. 51, 568-581 (2017). 2017:2398696 (2017). Calvi GP, Aud FF, Ferraz IDK, Pritchard HW, Kranner I. Analyses of several Egea J, Fabregat I, Frapart YM, Ghezzi P, Görlach A, Jansen-Dürr P, seed viability markers in individual recalcitrant seeds of Eugenia stipi- Daiber A. European contribution to the study of ROS: A summary of tata McVaugh with totipotent germination. Plant Biol. 19, 6-13 (2017). the findings and prospects for the future from the COST action BM1203 Buchner O, Roach T, Gertzen J, Schenk S, Karadar M, Stöggl W, Miller (EU-ROS). Redox Biol. 13, 94-162 (2017). R, Bertel C, Neuner G, Kranner I. Drought affects the heat-hardening

publications | Cavinato M, Jansen-Dürr P. Molecular mechanisms of UVB-induced capacity of alpine plants as indicated by changes in xanthophyll cycle senescence of dermal fibroblasts and its relevance for photoaging of pigments, singlet oxygen scavenging, α-tocopherol and plant hor- the human skin. Exp. Gerontol. 94, 78-82 (2017). mones. Environ. Exp. Bot. 133, 159-175 (2017). Erusalimsky JD, Grillari J, Grune T, Jansen-Duerr P, Lippi G, Sinclair AJ, García-Plazaola JI, Portillo-Estrada M, Fernández-Marín B, Kannaste Tegnér J, Viña J, Durrance-Bagale A, Miñambres R, Viegas M, Rodríguez- A, Niinemets U. Emissions of carotenoid cleavage products upon heat Mañas L. FRAILOMIC Consortium. In Search of ‘Omics’-Based Biomark- shock and mechanical wounding from a foliose lichen. Environ. Exp. ers to Predict Risk of Frailty and Its Consequences in Older Individuals: Bot. 133, 87-97 (2017). The FRAILOMIC Initiative. Gerontology 62(2), 182-90 (2016). Roach T, Baur T, Stöggl W, Krieger-Liszkay A. Chlamydomonas re- Lippi G, Jansen-Duerr P, Viña J, Durrance-Bagale A, Abugessaisa I, Go- inhardtii responding to high light: a role for 2-propenal (acrolein). mez-Cabrero D, Tegnér J, Grillari J, Erusalimsky J, Sinclair A, Rodriguez- Physiol. Plant. 161, 75-87 (2017). Manãs L. FRAILOMIC consortium.Laboratory biomarkers and frailty: Roach T, Na CS. LHCSR3 affects de-coupling and re-coupling of LHCII to presentation of the FRAILOMIC initiative. Clin. Chem. Lab. Med. 53(10), PSII during state transitions in Chlamydomonas reinhardtii. Sci. Rep. 7, e253-5 (2016). 43145 (2017). Galland M, He D, Lounifi I, Arc E, Clement G, Balzergue S, Huguet S, I. Kranner Cueff G, Godin B, Collet B, Granier F, Morin H, Tran J, Valot B, Rajjou L. Karadar M, Neuner G, Kranner I, Holzinger A, Buchner O. Solar irra- An integrated “Multi-Omics” Comparison of embryo and endosperm diation levels during simulated long and short-term heat waves sig- tissue-specific features and their impact on rice seed quality. Front. nificantly influence heat survival, pigment and ascorbate composition, Plant Sci. 8, 1984 (2017). and free radical scavenging activity in alpine Vaccinium gaultherioides. García-Plazaola JI, Fernández-Marín B, Ferrio JP, Alday JG, Hoch G, Physiol. Plantarum doi: 10.1111/ppl.12686 (2017). Landais D, Milcu A, Tissue DT, Voltas J, Gessler A, Roy J, de Dios VR. Fernández-Marín B, Míguez F, Méndez-Fernández L, Agut A, Becer- Endogenous circadian rhythms in pigment composition induce changes ril JM, García-Plazaola JI, Kranner I, Colville L. Seed carotenoid and in photochemical efficiency in plant canopies. Plant Cell Environ. 40, tocochromanol composition of wild Fabaceae species is shaped by 1153-1162 (2017). phylogeny and ecological factors. Front. Plant Sci. 8, 1428 (2017). Míguez F, Fernández-Marín B, Becerril JM, García-Plazaola JI. Diver- Calvi GP, Anjos AMG, Kranner I, Pritchard HW, Ferraz IDK. Exceptional sity of winter photoinhibitory responses: a case study in co-occurring flooding tolerance in the totipotent recalcitrant seeds of Eugenia stipi- lichens, mosses, herbs and woody plants from subalpine environments. tata. Seed Sci. Res. 27, 121-130 (2017). Physiol. Plantarum 160, 282-296 (2017). 72  Publications Publications  73

Widner FJ, Lawrence AD, Deery E, Heldt D, Frank S, Gruber K, Wurst K, Warren MJ, Kräutler B.Total Synthesis, Structure and Biological Activity of Adenosylrhodibalamin – the Nonnatural Rhodium Homologue of

Candotto Carniel F, Gerdol M, Montagner A, Banchi E, De Moro G, Coenzyme B12. Angew. Chem. Int. Ed. 55, 11281-11289 (2016). Manfrin C, Muggia L, Pallavicini A, Tretiach M. New features of desicca- Totalsynthese, Struktur und biologische Aktivität von Adenosylrhod- tion tolerance in the lichen photobiont Trebouxia gelatinosa revealed ibalamin, dem nichtnatürlichen Rhodiumhomologen von Coenzym B12. by a transcriptomic approach. Plant Mol. Biol. 91, 319-339 (2016). Angew. Chem. 128, 11451-11456 (2016). Eckert EM, Di Cesare A, Stenzel B, Fontaneto D, Corno G. Daphnia as a Erhart T, Mittelberger C, Vergeiner C, Scherzer G, Holzner B, Robatscher refuge for an antibiotic resistance gene in an experimental freshwater P, Oberhuber M, Kräutler B.Chlorophyll Catabolites in Senescent Leaves community. Sci. Total Environ. 571, 77-81 (2016). of the Plum Tree (Prunus domestica). Chem. & Biodiversity 13, 1441- Peguero-Pina JJ, Sisó S, Fernández-Marín B, Flexas J, Galmés J, 1453 (2016). García-Plazaola JI, Niinemets U, Sancho-Knapik D, Gil-Pelegrin E. Leaf Guzzo MB, Nguyen HT, Pham TH, Wyszczelska-Rokiel M, Jakubowski functional plasticity decreases the water consumption without further H, Wolff KA, Ogwang S, Timpona JL, Gogula S, Jacobs MR, Ruetz M, consequences for carbon uptake in Quercus coccifera L. under Mediter- Kräutler B, Jacobsen DW, Zhang GF, Nguyen L. Methylfolate Trap Pro- ranean conditions. Tree Physiol. 36, 356-367 (2016). motes Bacterial Thymineless Death by Sulfa Drugs. PLOS Pathogens 12, e1005949, (2016). B. Kräutler, T. Müller C Li, K Wurst, S Jockusch, K Gruber, M Podewitz, KR Liedl, B.Kräutler. Kräutler B. Breakdown of Chlorophyll in Higher Plants - Phyllobilins as Chlorophyll-Derived Yellow Phyllobilins of Higher Plants are Medium- Abundant, Yet Hardly Visible Signs of Ripening, Senescence and Cell Responsive, Chiral Photoswitches. Angew. Chem. Int. Ed. 55, 15760- Death (Review). Angew. Chem. Int. Ed. 55, 4882-4907 (2016). 15765, (2016). Der Chlorophyll-Abbau in höheren Pflanzen – Phyllobiline als weit- Von Chlorophyll abstammende gelbe Phyllobiline höherer Pflanzen verbreitete, aber kaum sichtbare Zeichen von Reifung, Seneszenz und

publications | als umgebungsgesteuerte, chirale Photoschalter. Angew. Chem. 128, Zelltod. Angew. Chem. 128, 4964-4990 (2016). 15992-15997 (2016). Netsomboon K, Feßler A, Erletz L, Prüfert F, Ruetz M, Kieninger C, Miller NA, Wiley TE, Spears KG, Ruetz M, Kieninger C, Krautler B,

Kräutler B, Bernkop-Schnürch A. Vitamin B12 and derivatives-in vitro Sension RJ. Toward the Design of Photoresponsive Conditional Antivi- permeation studies across Caco-2 cell monolayers and freshly excised tamins B12: A Transient Absorption Study of an Arylcobalamin and an rat intestinal mucosa. Int. J. Pharm. 497, 129-135 (2016). Alkynylcobalamin. J. Am. Chem. Soc. 138, 14250-14256 (2016). Li C, Kräutler B. Zn-complex of a natural yellow chlorophyll catabolite. Mireku SA, Ruetz MM, Zhou T, Korkhov VM, Kräutler B, Locher J. Porph. Phthalocyanines 20, 388-396 (2016).K Gruber, Kräutler B. KP.Conformational Change of a Tryptophane Residue in BtuF Facilitätes Coenzyme B Repurposed for Photo-regulation of Gene Expression. 12 Binding and Transport of Cobinamide by the Bacterial Vitamin B12 Angew. Chem. Int. Ed. 55, 5638-5640 (2016). Transporter BtuCD-F. Scient Reports 7, 41575 (2017).

Coenzym B12 – Umfunktioniert für die Photo-Regulation der Genex- Mittelberger C, Yalcinkaya H, Pichler C, Gasser J, Scherzer G, Erhart T, pression. Angew. Chem. 128, 5728-5730 (2016). Schumacher S, Holzner B, Janik K, Robatscher P, Muller T, Krautler B, Helliwell KE, Lawrence AD, Holzer A, Kudahl UJ, Sasso S, Kräutler B, Oberhuber M. Pathogen-Induced Leaf Chlorosis: Products of Chloro- Scanlan DJ, Warren MJ, Smith AG. Cyanobacteria and eukaryotic algae phyllBreakdown Found in Degreened Leaves of Phytoplasma-Infect-

use different chemical variants of vitamin B12. Current Biol. 26, 999-1008 edApple (Malus × domestica Borkh.) and Apricot (Prunus armeniaca L.) (2016). Trees Relate to the Pheophorbide a /Phyllobilin Pathway. J. Li C, Wurst K, Feng Y, Kräutler B. Synthesis, spectroscopic and crystal- Agr. Food Chem. 65, 2651-2660 (2017). lographic analysis of the Zn-complex of a di-(b,b”-sulfoleno) pyrin Ruetz M, Shanmuganathan A, Gherasim C, Karasik A, Salchner R, – Model for Zn-complexes of bilirubin and of phylloxanthobilins. Kieninger C, Wurst K, Banerjee R, Koutmos M, Kräutler B. Antivitamin Monatsh. Chem. 147, 1031-1036 (2016). B12 Inhibition of Human B12-Processing Enzyme CblC - Crystal Structure Banala S, Wurst K, Kräutler B. Panchromatic π-Extended Porphyrins of Abortive Ternary Complex with Cosubstrate Glutathione. Angew. from Conjugation with Quinones. ChemPlusChem. 81, 477-488 (2016). Chem. Int. Ed. 56, 7387-7392 (2017). Scherl M, Müller T, Kreutz CR, Huber RG, Zass E, Liedl KR, Kräutler B. Inhibierung des humanen B12-verarbeitenden Enzyms CblC durch An- Chlorophyll Catabolites in Fall Leaves of the Wych Elm Tree Present a tivitamine B12 – Kristallstruktur des inaktiven ternären Komplexes mit Novel Glycosylation Motif.Chem. Eur. J. 22, 9498-9503 (2016). dem Kosubstrat Glutathion. Angew. Chem. 129, 7493-7498 (2017) 74  Publications Publications  75

Stemeseder T, Freier R, Wildner S, Fuchs JE, Briza P, Lang R, Batanero E, Lidholm J, Liedl KR, Campo P. Crystal structure of Pla l 1 reveals both Li Z, Shanmuganathan A, Ruetz M, Yamada K, Lesniak NA, Kräutler structural similarity and allergenic divergence within the Ole e 1–like B, Brunold TC, Koutmos M Banerjee R. Glutathionyl-Cobalamin is an protein family. J. Allergy Cli. Immunol. 140(1), 277-80 (2017). Schmitt MK, Podewitz M, Liedl KR, Huppertz H. High-Pressure Synthesis Intermediate in Thiol Oxidation Catalyzed by the B12 Trafficking Protein CblC. J. Biol. Chem. 292, 9733-974 (2017). and Characterization of the Ammonium Yttrium Borate (NH4) YB8O14. Brenig CN, Ruetz M, Kieninger C, Wurst K, Kräutler B. Alpha- and Inorg. Chem. 56(22), 14291-9 (2017). beta-Diastereoisomers of Phenylcobalamin from Cobalt-Arylation with Schauperl M, Podewitz M, Ortner TS, Waibl F, Thoeny A, Loerting T, Liedl Diphenyliodonium Chloride. Chemistry – Europ. J. 23, 9726-9731 (2017). KR. Balance between hydration enthalpy and entropy is important for Banala S, Fokong S, Brand Ch, Andreou Ch, Kräutler B, Rueping M, ice binding surfaces in Antifreeze Proteins. Sci. Rep. 7(1), 11901 (2017). Kiessling F. Quinone-Fused Porphyrins as Contrast Agents for Photoa- Schauperl M, Czodrowski P, Fuchs JE, Huber RG, Waldner BJ, Podewitz coustic Imaging. Chem. Sci. 8, 6176-6181 (2017). M, Kramer C, Liedl KR. Binding Pose Flip Explained via Enthalpic and

Widner FJ, Gstrein F, Kräutler B. Partial Synthesis of Coenzyme B12 from Entropic Contributions. J. Cheml. Inf. Model. 57(2), 345-54 (2017). Cobyric Acid. Helv. Chim. Acta 100, e1700170 (2017). Rossi D, Rui M, Di Giacomo M, Schepmann D, Wuensch B, Monteleone Bloch JS, Ruetz M, Kräutler B, Locher KP.Structure of the human trans- S, Liedl KR, Collina S. Gaining in pan-affinity towards sigma 1 and sigma cobalamin beta domain in four distinct states. PloS One 12(9): e0184932 2 receptors. SAR studies on arylalkylamines. Bioorg. Med. Chem. 25(1), (2017). 11-9 (2017). Mutti E, Hunger M, Fedosov S, Nexo E, Kräutler B. Organometallic Peduto A, Scuotto M, Krauth V, Roviezzo F, Rossi A, Temml V, Esposito V,

DNA-B12-Conjugates as Potential Oligonucleotide Vectors - Synthesis, Stuppner H, Schuster D, D’Agostino B. Optimization of benzoquinone Structural and Binding Studies with Human Cobalamin-Transport Pro- and hydroquinone derivatives as potent inhibitors of human 5-lipoxy-

publications | teins. Chembiochem. 18, 2280-2291 (2017). genase. Eur. J. Med. Chem. 127, 715-26 (2017). Moser S, Scherzer G, Kräutler B. On the Nature of Isomeric Nonfluores- Obermoser V, Mauersberger R, Schuster D, Czifersky M, Lipova M, cent Chlorophyll Catabolites in Leaves and Fruit - A Study with a Ubiq- Siegl M, Kintscher U, Gust R. Importance of 5/6-aryl substitution on the uitous Phylloleucobilin and its Main Isomerization Product. Chemistry pharmacological profile of ʹ4 -((2-propyl-1H-benzo [d] imidazol-1-yl) and Biodiversity 14, e1700368 (2017). methyl)-[1, 1ʹ-biphenyl]-2-carboxylic acid derived PPARγ agonists. Eur. J. Kuprian E, Munkler C, Resnyak A, Zimmermann S, Tuong TD, Gierlinger Med. Chem. 126, 590-603 (2017). N, Müller T, Livingston III DP, Neuner G. Complex bud architecture and Monteleone S, Lieb A, Pinggera A, Negro G, Fuchs JE, Hofer F, Striessnig cell-specific chemical patterns enable supercooling of Picea abies bud J, Tuluc P, Liedl KR. Mechanisms Responsible for ω-Pore Currents in Ca v primordia. Plant Cell Environ. 40, 3101–3112 (2017). Calcium Channel Voltage-Sensing Domains. Biophys. J. 113(7), 1485-95 (2017). K. Liedl, D. Schuster Monteleone S, Fuchs JE, Liedl KR. Molecular Connectivity Predefines Vuorinen A, Engeli RT, Leugger S, Kreutz CR, Schuster D, Odermatt Polypharmacology: Aliphatic Rings, Chirality, and sp3 Centers Enhance A, Matuszczak B. Phenylbenzenesulfonates and-sulfonamides as Target Selectivity. Front. Pharmacol. 8, 552 (2017). 17β-hydroxysteroid dehydrogenase type 2 inhibitors: Synthesis and Hoffmann A, Richter M, von Grafenstein S, Walther E, Xu Z, Schumann SAR-analysis. Bioorg. Medl. Chem. Lett. 27(13), 2982-5 (2017). L, Grienke U, Mair CE, Kramer C, Rollinger JM. Discovery and Character- Vuorinen A, Engeli RT, Leugger S, Bachmann F, Akram M, Atanasov AG, ization of Diazenylaryl Sulfonic Acids as Inhibitors of Viral and Bacterial Waltenberger B, Temml V, Stuppner H, Krenn L. Potential Antiosteopo- Neuraminidases. Front. Microbiol. 8, 205 (2017). rotic Natural Product Lead Compounds That Inhibit 17β-Hydroxysteroid Hochleitner J, Akram M, Ueberall M, Davis RA, Waltenberger B, Dehydrogenase Type 2. J. Nat. Prod. 80(4), 965-74 (2017). Stuppner H, Sturm S, Ueberall F, Gostner JM, Schuster D. A combinato- Vitzthum D, Schauperl M, Liedl KR, Huppertz H. High-pressure synthesis rial approach for the discovery of cytochrome P450 2D6 inhibitors from and crystal structure of In3B5O12. Zeitschrift für Naturforschung B nature. Sci. Rep. 7(1), 8071 (2017). 72(1), 69-76 (2017). Hitzenberger M, Schuster D, Hofer TS. The Binding Mode of the Sonic Temml V, Garscha U, Romp E, Schubert G, Gerstmeier J, Kutil Z, Ma- Hedgehog Inhibitor Robotnikinin, a combined Docking and QM/MM tuszczak B, Waltenberger B, Stuppner H, Werz O. Discovery of the first MD Study. Front. Chem. 5, 76 (2017). dual inhibitor of the 5-lipoxygenase-activating protein and soluble Haq FU, Abro A, Raza S, Liedl KR, Azam SS. Molecular dynamics simula- epoxide hydrolase using pharmacophore-based virtual screening. Sci. tion studies of novel β-lactamase inhibitor. J. Mol. Graph. Model. 74, Rep. 7, 42751 (2017). 143-52 (2017). 76  Publications Publications  77

Zatelli GA, Temml V, Kutil Z, Landa P, Vanek T, Schuster D, Falkenberg M. Miconidin Acetate and Primin as Potent 5-Lipoxygenase Inhibitors Hanakova Z, Hosek J, Kutil Zf, Temml V, Landa Pe, Vaneǩ Ts, Schuster from Brazilian Eugenia hiemalis (Myrtaceae). Planta Medica Letters D, Dall’Acqua S, Cvackǎ J, Polanský Oe. Anti-inflammatory Activity of 3(01), e17-e9 (2016). Natural Geranylated Flavonoids: Cyclooxygenase and Lipoxygenase Xu Z, Von Grafenstein S, Walther E, Fuchs JE, Liedl KR, Sauerbrei A, Inhibitory Properties and Proteomic Analysis. J. Nat. Prod. 80(4), 999- Schmidtke M. Sequence diversity of NanA manifests in distinct enzyme 1006 (2017). kinetics and inhibitor susceptibility. Sci. Rep. 6, 25169 (2016). Grutsch S, Fuchs JE, Ahammer L, Kamenik AS, Liedl KR, Tollinger M. Wang Y, Gkeka P, Fuchs JE, Liedl KR, Cournia Z. DPPC-cholesterol Conformational Flexibility Differentiates Naturally Occurring Bet v 1 phase diagram using coarse-grained Molecular Dynamics simulations. Isoforms. Int. J. Mol. Sci. 18(6), 1192 (2017). Biochim. Biophys. Acta (BBA)-Biomembranes 1858(11), 2846-57 (2016). Garscha U, Romp E, Pace S, Rossi A, Temml V, Schuster D, König S, Gerst- Waltenberger B, Garscha U, Temml V, Liers J, Werz O, Schuster D, meier J, Liening S, Werner M. Pharmacological profile and efficiency in Stuppner H. Discovery of potent soluble epoxide hydrolase (sEH) inhibi- vivo of diflapolin, the first dual inhibitor of 5-lipoxygenase-activating tors by pharmacophore-based virtual screening. J. Chem. Inf. Model. protein and soluble epoxide hydrolase. Sci. Rep. 7(1), 9398 (2017). 56(4), 747-62 (2016). Engeli RT, Rohrer SR, Vuorinen A, Herdlinger S, Kaserer T, Leugger S, Waltenberger B, Atanasov AG, Heiss EH, Bernhard D, Rollinger JM, Schuster D, Odermatt A. Interference of Paraben Compounds with Breuss JM, Schuster D, Bauer R, Kopp B, Franz C. Drugs from nature Estrogen Metabolism by Inhibition of 17β-Hydroxysteroid Dehydroge- targeting inflammation (DNTI): a successful Austrian interdisciplinary nases. Int. J. Mol. Sci. 18(9), 2007 (2017). network project. Monatsh. Chem. 147(3), 479-91 (2016). E Fuchs J, Schilling O, R Liedl K. Determinants of Macromolecular Speci- Waldner BJ, Fuchs JE, Schauperl M, Kramer C, Liedl KR. Protease Inhibi- ficity from Proteomics-De rived Peptide Substrate Data. Curr. Protein. tors in View of Peptide Substrate Databases. J. Chem. Inf. Model. 56(6),

publications | Pept. Sci. 18(9), 905-13 (2017). 1228-35 (2016). Dreier D, Latkolik S, Rycek L, Schnürch M, Dymáková A, Atanasov AG, Waldner BJ, Fuchs JE, Huber RG, von Grafenstein S, Schauperl M, Ladurner A, Heiss EH, Stuppner H, Schuster D. Linked magnolol dimer Kramer C, Liedl KR. Quantitative Correlation of Conformational Bind- as a selective PPARγ agonist–Structure-based rational design, synthesis, ing Enthalpy with Substrate Specificity of Serine Proteases. J. Phys. and bioactivity evaluation. Sci. Rep. 7(1), 13002 (2017). Chem. B 120(2), 299-308 (2016). Bruns J, Podewitz M, Schauperl M, Joachim B, Liedl KR, Huppertz H. Scherl M, Müller T, Kreutz CR, Huber RG, Zass E, Liedl KR, Kräutler B. CaB2S4O16: A Borosulfate Exhibiting a New Structure Type with Phyl- Chlorophyll Catabolites in Fall Leaves of the Wych Elm Tree Present a losilicate Analogue Topology. Chemistry 23(66), 16773-81 (2017). Novel Glycosylation Motif. Chemistry 22(28), 9498-503 (2016). Bernard J, Köck E-M, Huber RG, Liedl KR, Call L, Schlögl R, Grothe H, Schauperl M, Podewitz M, Waldner BJ, Liedl KR. Enthalpic and Entropic Loerting T. Carbonic acid monoethyl ester as a pure solid and its confor- Contributions to Hydrophobicity. J. Chem. Theory Comput. 12(9), 4600- mational isomerism in the gas-phase. Rsc Adv. 7(36), 22222-33 (2017). 10 (2016). Beck KR, Kaserer T, Schuster D, Odermatt A. Virtual screening appli- Scharinger B, Messner B, Türkcan A, Schuster D, Vuorinen A, Pitterl F, cations in short-chain dehydrogenase/reductase research. J. Steroid Heinz K, Arnhard K, Laufer G, Grimm M. Leoligin, the major lignan Biochem. Mol. Biol. 171, 157-177 (2017). from Edelweiss, inhibits 3-hydroxy-3-methyl-glutaryl-CoA reductase Beck KR, Bächler M, Vuorinen A, Wagner S, Akram M, Griesser and reduces cholesterol levels in ApoE−/− mice. J. Mol. Cell. Cardiol. 99, U, Temml V, Klusonova P, Yamaguchi H, Schuster D. Inhibition of 35-46 (2016). 11β-hydroxysteroid dehydrogenase 2 by the fungicides itraconazole Schaible AM, Filosa R, Krauth V, Temml V, Pace S, Garscha U, Liening and posaconazole. Biochem. Pharmacol. 130, 93-103 (2017). S, Weinigel C, Rummler S, Schieferdecker S. The 5-lipoxygenase inhibi- Akram M, Waratchareeyakul W, Haupenthal J, Hartmann RW, Schuster tor RF-22c potently suppresses leukotriene biosynthesis in cellulo and D. Pharmacophore modeling and in silico/in vitro screening for human blocks bronchoconstriction and inflammation in vivo. Biochem. Phar- cytochrome P450 11B1 & cytochrome P450 11B2 inhibitors. Front. macol. 112, 60-71 (2016). Chem. 5, 104 (2017). Reintjes A, Fuchs JE, Kremser L, Lindner HH, Liedl KR, Huber LA, Valovka Ahammer L, Grutsch S, Kamenik AS, Liedl KR, Tollinger M. Structure of T. Asymmetric arginine dimethylation of RelA provides a repressive the Major Apple Allergen Mal d 1. J. Agric. Food Chem. 65(8), 1606-12 mark to modulate TNFα/NF-κB response. Proc. Natl. Acad. Sci. USA (2017). 113(16), 4326-31 (2016). 78  Publications Publications  79

Machado Y, Freier R, Scheiblhofer S, Thalhamer T, Mayr M, Briza P, Abbasi S, Raza S, Azam SS, Liedl KR, Fuchs JE. Interaction mechanisms Grutsch S, Ahammer L, Fuchs JE, Wallnoefer HG. Fold stability during of a melatonergic inhibitor in the melatonin synthesis pathway. J. Mol. endolysosomal acidification is a key factor for allergenicity and immu- Liq. 221, 507-17 (2016). nogenicity of the major birch pollen allergen. J. Allergy. Clin. Immunol. 137(5), 1525-34 (2016). D. Meyer, P. Aanstad Loeffler JR, Ehmki ES, Fuchs JE, Liedl KR. Kinetic barriers in the isom- Facchinello N, Tarifeno-Saldivia E, Grisan E, Schiavone M, Peron M, erization of substituted ureas: implications for computer-aided drug Mongera A, Ek O, Schmitner N, Meyer D, Peers B, Tiso N, Argenton design. J. Comput. Aided Mol. Des. 30(5), 391-400 (2016). F. Tcf7l2 plays pleiotropic roles in the control of glucose homeostasis, Li C, Wurst K, Jockusch S, Gruber K, Podewitz M, Liedl KR, Kräutler B. pancreas morphology, vascularization and regeneration. Sci. Rep. 7, Chlorophyll-Derived Yellow Phyllobilins of Higher Plants as Medium- 9605 (2017). Responsive Chiral Photoswitches. Angew. Chem. Int. Ed. Engl. 55(51), Li J, Casteels T, Frogne T, Ingvorsen C, Honore C, Courtney M, Huber 15760-5 (2016). KV, Schmitner N, Kimmel RA, Romanov RA, Sturtzel C, Lardeau CH, Kratz JM, Mair CE, Oettl SK, Saxena P, Scheel O, Schuster D, Hering S, Klughammer J, Farlik M, Sdelci S, Vieira A, Avolio F, Briand F, Baburin I, Rollinger JM. hERG Channel Blocking Ipecac Alkaloids Identified by Majek P, Pauler FM, Penz T, Stukalov A, Gridling M, Parapatics K, Bar- Combined In Silico–In Vitro Screening. Planta Med. 82(11/12), 1009-15 bieux C, Berishvili E, Spittler A, Colinge J, Bennett KL, Hering S, Sulpice (2016). T, Bock C, Distel M, Harkany T, Meyer D, Superti-Furga G, Collombat P, Kramer C, Mochalski P, Unterkofler K, Agapiou A, Ruzsanyi V, Liedl KR. Hecksher-Sorensen J, Kubicek S. Artemisinins. Target GABAA Receptor Prediction of blood: air and fat: air partition coefficients of volatile or- Signaling and Impair alpha Cell Identity. Cell 168, 86-100 e115 (2017).

publications | ganic compounds for the interpretation of data in breath gas analysis. Schmitner N, Kohno K, Meyer D. ptf1a+ , ela3l- cells are developmen- J. Breath Res. 10(1), 017103 (2016). tally maintained progenitors for exocrine regeneration following Kaserer T, Rigo R, Schuster P, Alcaro S, Sissi C, Schuster D. Optimized virtual extreme loss of acinar cells in zebrafish larvae. Dis. Model. Mech. 10, screening workflow for the identification of novel G-quadruplex ligands. 307-321 (2017). Journal of chemical information and modeling 56(3), 484-500 (2016). Bachmann VA, Mayrhofer JE, Ilouz R, Tschaikner P, Raffeiner P, Röck R, Kaserer T, Obermoser V, Weninger A, Gust R, Schuster D. Evaluation Courcelles M, Apelt F, Lu TW, Baillie GS, Thibault P, Aanstad P, Stelzl U, of selected 3D virtual screening tools for the prospective identification Taylor SS, Stefan E. Gpr161 anchoring of PKA consolidates GPCR and of peroxisome proliferator-activated receptor (PPAR) γ partial agonists. cAMP signaling. Proc. Natl. Acad. Sci. 113, 7786 (2016). Europ. J. Med. Chem. 124, 49-62 (2016). Kimmel RA, Meyer D. Zebrafish pancreas as a model for development Kaserer T, Lantero A, Schmidhammer H, Spetea M, Schuster D. μ Opi- and disease. Methods Cell Biol. 134, 431-61 (2016). oid receptor: novel antagonists and structural modeling. Sci. Rep. 6, Ott E, Wendik B, Srivastava M, Pacho F, Töchterle S, Salvenmoser W, srep21548 (2016). Meyer D. Pronephric tubule morphogenesis in zebrafish depends on Kamenik AS, Kahler U, Fuchs JE, Liedl KR. Localization of Millisecond Mnx mediated repression of irx1b within the intermediate mesoderm. Dynamics: Dihedral Entropy from Accelerated MD. J. Chem. Theory Dev. Biol. 411(1), 101-14 (2016). Comput. 12(8), 3449-55 (2016). Glätzle M, Schauperl M, Hejny C, Tribus M, Liedl KR, Huppertz H. Or- R. Micura thorhombic HP-REOF (RE= Pr, Nd, Sm–Gd)–High-Pressure Syntheses and Neuner S, Falschlunger C, Fuchs E, Himmelstoss M, Ren A, Patel DJ, Single-Crystal Structures (RE= Nd, Sm, Eu). Zeitschrift für anorganische Micura R. Atom-Specific Mutagenesis Reveals Structural and Catalytic und allgemeine Chemie 2016(20), 1134-42 (2016). Roles for an Active-Site Adenosine and Hydrated Mg2+ in Pistol Ribo- Beck KR, Sommer TJ, Schuster D, Odermatt A. Evaluation of tetrabro- zymes. Angew. Chem. Int. Ed. Engl. 56, 15954-15958 (2017). mobisphenol A effects on human glucocorticoid and androgen recep- Zheng L, Mairhofer E, Teplova M, Zhang Y, Ma J, Patel DJ, Micura R, Ren tors: A comparison of results from human-with yeast-based in vitro A. Structure-based insights into self-cleavage by a four-way junctional assays. Toxicology 370, 70-7 (2016). twister-sister ribozyme. Nat. Commun. 8, 1180 (2017). Alsabil K, Suor-Cherer S, Koeberle A, Viault G, Lavaud A, Temml V, Riml C, Amort T, Rieder D, Gasser C, Lusser A, Micura R. Osmium-Medi- Waltenberger B, Schuster D, Litaudon M, Lorkowski S. Semisynthetic ated Transformation of 4-Thiouridine to Cytidine as Key To Study RNA and Natural Garcinoic Acid Isoforms as New mPGES-1 Inhibitors. Planta Dynamics by Sequencing. Angew. Chem. Int. Ed. Engl. 56, 13479-13483 Med. 82(11/12), 1110-6 (2016). (2017). 80  Publications Publications  81

Vušurović N, Altman RB, Terry DS, Micura R, Blanchard SC. Pseudoknot Formation Seeds the Twister Ribozyme Cleavage Reaction Coordinate. Ren A, Vušurović N, Gebetsberger J, Gao P, Juen M, Kreutz C, Micura J. Am. Chem. Soc. 139, 8186-8193 (2017). R, Patel DJ. Pistol ribozyme adopts a pseudoknot fold facilitating site- Danhart EM, Bakhtina M, Cantara WA, Kuzmishin AB, Ma X, Sanford specific in-line cleavage. Nat. Chem. Biol. 12, 702-708 (2016). BL, Košutić M, Goto Y, Suga H, Nakanishi K, Micura R, Foster MP, Musier- Melnikov S, Mailliot J, Shin BS, Rigger L, Yusupova G, Micura R, Dever Forsyth K. Conformational and chemical selection by a trans-acting TE, Yusupov M. Crystal Structure of Hypusine-Containing Translation editing domain. Proc. Natl. Acad. Sci. USA 114, E6774-E6783 (2017). Factor eIF5A Bound to a Rotated Eukaryotic Ribosome. J. Mol. Biol. 428, Ren A, Micura R, Patel DJ. Structure-based mechanistic insights into 3570-3576 (2016). catalysis by small self-cleaving ribozymes. Curr. Opin. Chem. Biol. 41, Flür S, Micura R. Chemical synthesis of RNA with site-specific meth- 71-83 (2017). ylphosphonate modifications. Methods 107, 79-88 (2016). Riml C, Lusser A, Ennifar E, Micura R. Synthesis, Thermodynamic Frener M, Micura R. Conformational Rearrangements of Individual Properties, and Crystal Structure of RNA Oligonucleotides Containing Nucleotides during RNA-Ligand Binding Are Rate-Differentiated. J. Am. 5-Hydroxymethylcytosine. J. Org. Chem. 82, 7939-7945 (2017). Chem. Soc. 138, 3627-3630 (2016). Glasner H, Riml C, Micura R, Breuker K. Label-free, direct localization and relative quantitation of the RNA nucleobase methylations m6A, B. Pelster, A. Sandbichler, T. Schwerte m5C, m3U, and m5U by top-down mass spectrometry. Nucleic Acids Res. Wood CM, Pelster B, Giacomin M, Sadauskas H, Almeida-Val VF, Val AL. 45, 8014-8025 (2017). The transition from water-breathing to air-breathing is associated with Riml C, Micura R. Automated Chemical Solid-Phase Synthesis and De- a shift in ion uptake from gills to gut: a study of two closely-related ery- thrinid teleosts, Hoplerythrinus unitaeniatus and Hoplias malabaricus. publications | protection of 5-Hydroxy­methylcytosine-Containing RNA. Methods Mol. Biol. 1562, 295-302 (2017). J. Comp. Physiol. B 186, 431-445 (2016). Mairhofer E, Fuchs E, Micura R. Facile synthesis of a 3-deazaadenosine Pelster B, Schneebauer G, Dirks RP. Anguillicola crassus infection signifi- phosphoramidite for RNA solid-phase synthesis. Beilstein J. Org. Chem. cantly affects the silvering related modifications in steady state mRNA 12, 2556-2562 (2016). levels in gas gland tissue of the European eel. Frontiers in Physiology Neuner S, Kreutz C, Micura R. The synthesis of 15N(7)-Hoogsteen face- 7, 175 (2016). labeled adenosine phosphoramidite for solid-phase RNA synthesis. Pelster B, Giacomin M, Wood CM, Val AL. Improved ROS defence in Monatsh. Chem. 148, 149-155 (2017). the swimbladder of a facultative air-breathing erythrinid fish, jeju, Amort T, Rieder D, Wille A, Khokhlova-Cubberley D, Riml C, Trixl L, Jia compared to a non-air-breathing close relative, traira. J. Comp. Physiol. XY, Micura R, Lusser A. Distinct 5-methylcytosine profiles in poly(A) RNA B. 186, 615-624 (2016). from mouse embryonic stem cells and brain. Genome Biol. 18, 1 (2017). Schneebauer G, Hanel R, Pelster B. Anguillicola crassus impairs the Gebetsberger J, Micura R. Unwinding the twister ribozyme: from struc- silvering related enhancements of the ROS defense capacity in swim- ture to mechanism. Wiley Interdiscip. Rev. RNA. 8, e1402, doi: 10.1002/ bladder tissue of the European eel (Anguilla anguilla). J. Comp. Physiol. wrna.1402 (2017). B. 186, 867-877 (2016). Jud L, Micura R. An Unconventional Acid-Labile Nucleobase Protection Kwong R, Kumai Y, Tzaneva V, Azzi E, Hochhold N, Robertson C, Pelster Concept for Guanosine Phosphoramidites in RNA Solid-Phase Synthesis. B, Perry SF. Inhibition of calcium uptake during hypoxia in developing Chemistry 23, 3406-3413 (2016). zebrafish, Danio rerio, is mediated by hypoxia-inducible factor. J. Exp. Melnikov S, Mailliot J, Rigger L, Neuner S, Shin BS, Yusupova G, Dever Biol. 219, 3988-3995 (2016). TE, Micura R, Yusupov M. Molecular insights into protein synthesis with Sandbichler AM, Höckner M. Cadmium Protection Strategies-A Hidden proline residues. EMBO Rep. 17, 1776-1784 (2016). Trade-Off? Int. J. Molec. Sci. 17/1, No. E139. (2016). Sothiselvam S, Neuner S, Rigger L, Klepacki D, Micura R, Vázquez-Laslop Piechnik CA, Höckner M, de Souza MR, Donatti L, Tomanek L. Time N, Mankin AS. Binding of Macrolide Antibiotics Leads to Ribosomal course of lead induced proteomic changes in gill of the Antarctic limpet Selection against Specific Substrates Based on Their Charge and Size. Nacella concinna (Gastropoda: Patellidae). J. Proteomics 151, 145-161 Cell Rep. 16, 1789-1799 (2016). (2016). Riml C, Micura R. Synthesis of 5-Hydroxymethylcytidine- and 5-Hydroxy- Jonz, MG, Buck LT, Perry SF, Schwerte T, Zaccone G. Sensing and surviv- methyl-uridine-Modified RNA. Synthesis 48, 1108-1116 (2016). ing hypoxia in vertebrates. Ann. NY Acad. Sci. 1365, 43-58 (2016). 82  Publications Publications  83

Noha SM, Schmidhammer H, Spetea M. Molecular docking, molecular Schindler RFR, Scotton C, Zhang J, Passarelli C, Ortiz-Bonnin B, Simrick S, dynamics and structure-activity relationship explorations of 14-oxygen- Schwerte T, Poon KL, Fang M, Rinné S, Froese A, Nikolaev VO, Grunert ated N-methylmorphinan-6-ones as potent µ-opioid receptor agonists. C, Müller T, Tasca G, Sarathchandra P, Drago F, Dallapiccola B, Rapezzi ACS Chem. Neurosci. 8, 1327-1337 (2017). C, Arbustini E, Di Raimo FR, Neri M, Selvatici R, Gualandi F, Fattori F, Spetea M, Eans SO, Ganno ML, Lantero A, Mairegger M, Toll L, Schmid- Pietrangelo A, Li W, Jiang H, Xu X, Bertini E, Decher N, Wang J, Brand T, hammer H, McLaughlin JP. Selective κ opioid receptor partial agonist Ferlini A. POPDC1S201F causes muscular dystrophy and arrhythmia by HS666 produces potent antinociception without inducing aversion af- affecting protein trafficking. J. Clinic. Invest. 1365, 43-58 (2016). ter i.c.v. administration in mice. Br. J. Pharmacol. 174, 2444-2456 (2017). Zobl S, Salvenmoser W, Schwerte T, Gebeshuber I, Schreiner M. Morpho Erli F, Guerrieri E, Ben Haddou T, Lantero A, Mairegger M, Schmidham- peleides butterfly wing imprints as structural colour stamp.Bioinspir. mer H, Spetea M. Highly potent and selective new diphenethylamines Biomim. 11(1), 016006 (2016). interacting with the κ-opioid receptor: Synthesis, pharmacology, and Jansen H, Liem M, Jong-Raadsen S, Dufour S, Weltzien FA, Swinkels W, structure-activity relationships. J. Med. Chem. 60, 579-7590 (2017). Koelewijn A, Palstra A, Pelster B, Spaink H, van den Thillart G, Dirks R, Dumitrascuta M, Ben Haddou T, Guerrieri E, Noha SM, Schläfer L, Henkel C. Rapid de novo assembly of the European eel genome from Schmidhammer H, Spetea M. Synthesis, pharmacology and molecular nanopore sequencing reads. Sci. Rep. 7, 7213 (2017). modeling studies on 6-desoxo-N-methylmorphinans as potent µ-opioid Schneebauer G, Dirks RP, Pelster B. Anguillicola crassus infection affects receptor agonists. J. Med. Chem. 60, 9407-9412 (2017). mRNA expression levels in gas gland tissue of European yellow and Schmidhammer H, Spetea M, Guerrieri E. Diphenethylamine derivatives silver eel. Plos One 12: e0183128 (2017). which are inter alia useful as analgesics and method for their produc- Drechsel V, Schauer K, Šrut M, Höckner M. Regulatory Plasticity of tion. United States Patent Application, US15/561,614 (2017).

publications | Earthworm wMT-2 Gene Expression. Int. J. Mol. Sci. 18, 1113 (2017). Schmidhammer H, Spetea M, Guerrieri E. Diphenethylamine derivatives Šrut M, Drechsel V, Höckner M. Low levels of Cd induce persisting which are inter alia useful as analgesics and method for their produc- epigenetic modifications and acclimation mechanisms in the earth- tion. European Patent Application, EP16712338.9 (2017). worm Lumbricus terrestris. PLoS One. 12(4), e0176047 (2017). R. Schneider, B. Auer H. Schmidhammer, M. Spetea Schweiger S, Matthes F, Posey K, Kickstein E, Weber S, Hettich MM, Kaserer T, Lantero A, Schmidhammer H, Spetea M, Schuster D. µ Opioid Pfurtscheller S, Ehninger D, Schneider R, Krauß S. Resveratrol induces receptor: Novel antagonists and structural modeling. Sci. Rep. 6, 21548 dephosphorylation of Tau by interfering with the MID1-PP2A complex. (2016). Sci. Rep. 7, 13753 (2017). Martin C, Oyen E, Mangelschots J, Bibian M, Ben Haddou T, Andrade Schindler S, Missbichler B, Walther C, Sponring M, Cserjan-Puschmann J, Gardiner J, Van Mele B, Madder A, Hoogenboom R, Spetea M, Ballet M, Auer B, Schneider R, Dürauer A. Npro fusion technology: On-column S. Injectable peptide hydrogels for controlled-release of opioids. Med. complementation to improve efficiency in biopharmaceutical produc- Chem. Comm. 7, 542-549 (2016). tion. Protein Expr. Purif. 120, 42-50 (2016). Guerrieri E, Bermudez M, Wolber G, Berzetei-Gurske IP, Schmidhammer Lemmermeyer T, Lamp B, Schneider R, Ziebuhr J, Tekes G, Thiel H-J. H, Spetea M. Structural determinants of diphenethylamines for interac- Characterization of monoclonal antibodies against feline coronavirus tion with the κ opioid receptor: Synthesis, pharmacology and molecular accessory protein 7b. Vet. Microbiol. 184, 11-19 (2016). modeling studies. Bioorg. Med. Chem. Lett. 26, 4769-4774 (2016). Lagard C, Chevillard L, Guillemyn K, Risède P, Laplanche J-L, Spetea J. Striessnig, N. Singewald, A. Koschak M, Ballet S, Mégarbane B. Bifunctional peptide-based opioid agonist/ Adori C, Barde S, Vas S, Ebner K, Su J, Svensson C, Mathé AA, Singewald nociceptin antagonist ligand for dual treatment of nociceptive and N, Reinscheid RR, Uhlén M, Kultima K, Bagdy G, Hökfelt T. Exploring neuropathic pain. Pain 158, 505-515 (2017). the role of neuropeptide S in the regulation of arousal: a functional Martin C, Oyen E, Van Wanseele Y, Ben Haddou T, Schmidhammer H, anatomical study. Brain Struct. Funct. 221, 3521-3546 (2016). Andrade J, Waddington L, Van Eeckhaut A, Van Mele B, Gardiner J, Mesirca P, Bidaud I, Briec F, Evain S, Torrente AG, Le Quang K, Mangoni Hoogenboom R, Madder A, Spetea M, Ballet S. Injectable peptide-based ME. G protein-gated IKACh channels as therapeutic targets for treat- hydrogel formulations for the extended in vivo release of opioids. Mat. ment of sick sinus syndrome and heart block. Proc. Natl. Acad. Sci. USA Today Chem. 3, 49-59 (2017). 113, 932-941 (2016). 84  Publications Publications  85

Martínez-Rivera A, Hao J, Tropea TF, Giordano TP, Kosovsky M, Rice RC, Lee A, Huganir RL, Striessnig J, Addy NA, Han S, Rajadhyaksha AM. En- Puschban Z, Sah A, Grutsch I, Singewald N, Dechant G. Reduced Anx- hancing VTA Cav13 L-type Ca2+ channel activity promotes cocaine and iety-Like Behavior and Altered Hippocampal Morphology in Female mood-related behaviors via overlapping AMPA receptor mechanisms in p75NTR(exon IV-/-) Mice. Front. Behav. Neurosci. 10, 103 (2016). the nucleus accumbens. Mol. Psychiatry 22, 1735-1745 (2017). Sartori SB, Maurer V, Murphy C, Schmuckermair C, Muigg P, Neu- Mastrolia V, Flucher SM, Obermair GJ, Drach M, Hofer H, Renström E, mann ID, Whittle N, Singewald N. Combined neuropeptide S and Schwartz A, Striessnig J, Flucher BE, Tuluc P. Loss of α2δ-1 Calcium Chan- D-cycloserine augmentation prevents the return of fear in extinction nel Subunit Function Increases the Susceptibility for Diabetes. Diabetes impaired rodents: advantage of dual vs. single drug approaches. Int. J. 66, 867-907 (2017). Neuropsychopharmacol. 19, 1-11 (2016). Monteleone Stefania, Lieb Andreas, Pinggera Alexandra, Negro Giulia, Schaufler J, Ronovsky M, Savalli G, Cabatic M, Sartori SB, Singewald N, Fuchs Julian E, Hofer Florian, Striessnig Jörg, Tuluc Petronel, Liedl Klaus Pollak DD. Fluoxetine normalizes disrupted light-induced entrainment R. Mechanisms Responsible for ω-Pore Currents in Cav Calcium Channel fragmented ultradian rhythms and altered hippocampal clock gene Voltage-Sensing Domains. Biophys. J. 113, 1485-1495 (2017). expression in an animal model of high trait anxiety- and depression- Ortner NJ, Bock G, Dougalis A, Kharitonova M, Duda J, Hess S, Tuluc T, related behavior. Ann. Med. 48, 17-27 (2016). Pomberger T, Stefanova N, Pitterl F, Ciossek T, Oberacher H, Draheim Stanika R, Campiglio M, Pinggera A, Lee A, Striessnig J, Flucher BE, HJ, Kloppenburg P, Liss B, Striessnig J. Lower Affinity of Isradipine for Obermair GJ. Splice variants of the Cav1.3 L-type calcium channel regu- L-Type Ca2+ Channels during Substantia Nigra Dopamine Neuron-like late dendritic spine morphology. Sci. Rep. 6, 34528 (2016). Activity: Implications for Neuroprotection in Parkinson’s Disease. J. Sultana N, Dienes B, Benedetti A, Tuluc P, Szentesi P, Sztretye M, Rainer Neurosci. 37, 6761-6777 (2017). J, Hess MW, Schwarzer C, Obermair GJ, Csernoch L, Flucher BE. Restrict- Pinggera A, Mackenroth L, Rump A, Schallner J, Beleggia F, Wollnik B,

publications | ing calcium currents is required for correct fiber type specification in Striessnig J. New Gain-of-Function Mutation Shows CACNA1D as Recur- skeletal muscle. Development 143, 1547-1559 (2016). rently Mutated Gene in Autism Spectrum Disorders and Epilepsy. Hum. Torrente AG, Mesirca P, Neco P, Rizzetto R, Dubel S, Barrere C, Sinneg- Mol. Genet. 26, 2923-2932 (2017). ger-Brauns M, Striessnig J, Richard S, Nargeot J, Gomez AM, Mangoni Tan GC, Negro G, Pinggera A, Tizen Laim NMS, Rose I, Ceral J, Ryska ME. L-type Cav1.3 Channels Regulate Ryanodine receptor-dependent A, Chin LK, Kamaruddin NA, Mohd Mokhtar N, A Jamal AR, Sukor N, calcium release during sino-atrial node pacemaker activity. Cardiovasc. Solar M, Striessnig J, Brown MJ, Azizan EA. Aldosterone-Producing Res. 109, 451-61 (2016). Adenomas: Histopathology-Genotype Correlation and Identification of Tuluc P, Benedetti B, Coste de Bagneaux P, Grabner M, Flucher BE. Two a Novel CACNA1D Mutation. Hypertension 70, 129-136 (2017). distinct voltage-sensing domains control voltage sensitivity and kinetics Toyoda F, Mesirca P, Dubel S, Ding W-G, Striessnig J, Mangoni ME, of current activation in CaV1.1 calcium channels. J. Gen. Physiol. 147, Matsuura H. CaV13 L-type Ca2+ channel contributes to the heartbeat 437 (2016). by generating a dihydropyridine-sensitive persistent Na+ current. Sci. Tuluc P, Yarov-Yarovoy V, Benedetti B, Flucher BE. Molecular Interac- Rep. 7, 7869 (2017). tions in the Voltage Sensor Controlling Gating Properties of CaV Alexander SP, Striessnig J, Kelly E, Marrion NV, Peters JA, Faccenda Calcium Channels. Structure 24, 261-271 (2016). E, Harding SD, Pawson AJ, Sharman JL, Southan C, Davies JA; CGTP Whittle N, Maurer V, Murphy C, Rainer J, Bindreither D, Hauschild M, Collaborators. THE CONCISE GUIDE TO PHARMACOLOGY 2017/18: Scharinger A, Oberhauser M, Keil T, Brehm C, Valovka T, Striessnig J, Voltage-gated ion channels. Br. J. Pharmacol. 174 (Suppl 1) S160-S194 Singewald N. Enhancing dopaminergic signaling and histone acetyla- (2017). tion promotes long-term rescue of deficient fear extinction. Transl. Alexander SP, Kelly E, Marrion NV, Peters JA, Faccenda E, Harding SD, Psychiatry 6, e974 (2016). Pawson AJ, Sharman JL, Southan C, Buneman OP, Cidlowski JA, Christo- Wille A, Amort T, Singewald N, Sartori SB, Lusser A. Dysregulation of se- poulos A, Davenport AP, Fabbro D, Spedding M, Striessnig J, Davies JA; lect ATP-dependent chromatin remodeling factors in high trait anxiety. CGTP Collaborators.THE CONCISE GUIDE TO PHARMACOLOGY 2017/18: Behav. Brain Res. 311, 141-146 (2016). Overview. Br. J. Pharmacol. 174 (Suppl 1) S1-S16 (2017). Härtner L, Keil TWM, Kreuzer M, Fritz EM, Wenning GK, Stefanova N, Ortner NJ, Striessnig J. L-type calcium channels as drug targets in CNS Fenzl T. Distinct Parameters in the EEG of the PLP α-SYN Mouse Model disorders. Channels (Austin) 10, 7-13 (2016). for Multiple System Atrophy Reinforce FaceValidity. Front. Behav. Neu- Pinggera A, Striessnig J. Cav 1.3 (CACNA1D) L-type Ca2+ channel dys- rosci. 10, 252 (2017). function in CNS disorders. J. Physiol. 594, 5839-5849 (2016). 86  Publications Publications  87

Striessnig J. Voltage-gated calcium channels - from basic mechanisms to Mailainer C, Schachner D, Sangiovanni E, Atanasov AG, Schwaiger S, disease. J. Physiol. 594, 5817-5821 (2016). Stuppner H, Heiss EH, Dirsch VM. Eurycomalactone Inhibits Expression Sartori BS, Singewald N. Neue pharmakologische Strategien zur Aug- of Endothelial Adhesion Molecules at a Post-Transcriptional Level. J. mentation von Extinktionslernen in der Angsttherapie. Neuroforum Nat. Prod. 80, 3186-3193 (2017). 23, 197-211 (2017). Marzocco S, Adesso S, Alilou M, Stuppner H, Schwaiger S. Anti-Inflam- Sartori BS, Singewald N. New pharmacological strategies for augment- matory and Anti-Oxidant Potential of the Root Extract and Constituents ing extinction learning in anxiety disorders. Neuroforum 23, A145-A156 of Doronicum austriacum. Molecules 22, 1003 (2017). (2017). Murauer A, Bakry R, Schottenberger H, Huck CW, Ganzera M. An in- Seitter H, Koschak A. Relevance of tissue specific subunit expression in novative monolithic zwitterionic stationary phase for the separation of channelopathies. Neuropharmacology 17, 30302-7 (2017). phenolic acids in coffee bean extracts by capillary electrochromatogra- Ebner K, Singewald N. Individual differences in stress susceptibility and phy. Anal. Chim. Acta 963, 136-142 (2017). stress inhibitory mechanisms. Current Opinion in Behavioral Sciences Murauer A, Ganzera M. Quantitative Determination of Lactones in 14, 54-64 (2017). Piper methysticum (Kava-Kava) by Supercritical Fluid Chromatography. Murphy CP, Singewald N. Potential of microRNAs as novel targets in Planta Med. 83, 1053-1057 (2017). the alleviation of pathological fear. Genes Brain Behav, doi: 10.1111/ Nebieridze VG, Skhirtladze AV, Kemertelidze EP, Ganzera M. Nucleo- gbb.12427. [Epub ahead of print] sides from Tribulus terrestris. Chemistry of Natural Compounds 53, Striessnig, Jörg: Editorial Board - Channels, 01.01.2007 lfd. 1010-1011 (2017). Singewald, Nicolas: Editor - Amino Acids, 01.01.1999 lfd. Nicolaus C, Junghanns S, Hartmann A, Murillo R, Ganzera M, Merfort I.

publications | In vitro studies to evaluate the wound healing properties of Calendula H. Stuppner, M. Ganzera officinalis extracts. J. Etnopharmacol. 196, 94-103 (2017). Dreier D, Latkolik S, Rycek L, Schnürch M, Dymáková A, Atanasov AG, Pataczek L, Cheilari A, Zikeli S, Sturm S, Stuppner H, Gruber S. Cen- Ladurner A, Heiss EH, Stuppner H, Schuster D, Mihovilovic MD, Dirsch taurium erythraea Cultivation Method for Optimal Yield and Product VM. Linked magnolol dimer as a selective PPARγ agonist – Structure- Quality. Journal of Herbs, Spices & Medicinal Plants 23, 193-215 (2017). based rational design, synthesis, and bioactivity evaluation. Sci. Rep. 7, Peduto A, Scuotto M, Krauth V, Roviezzo F, Rossi A, Temml V, Esposito V, 13002 (2017). Stuppner H, Schuster D, D´Agostino B., Schiraldi C, De Rosa M, Werz O, Gvazava L, Gorgaslidze N, Ganzera M, Skhirtladze A. A new lupane Filosa R. Optimization of benzoquinone and hydroquinone derivatives triterpene glycoside from Euphorbia boissierana Prokh. Trends in Phy- as potent inhibitors of human 5-lipoxygenase. Eur. J. Med. Chem. 127, tochemical Research 1, 149-152 (2017). 715-726 (2017). Haller J, Schwaiger S, Stuppner H, Gafner F, Ganzera M. Isolation of Schäfer S, Schwaiger S, Stuppner H. Aristolic Acid Derivates from the Three Triterpene Saponins, Including Two New Oleanane Derivatives, Bark of Antidesma ghaesembilla. Planta Med. 83, 1097-1102 (2017). from Soldanella alpina and Hydrophilic Interaction Liquid Chromatog- Siewert B, Langerman M, Hontani Y, Kennis JTM, Van Rixel VHS, Lim- raphy-Evaporative Light Scattering Detection of these Three Saponins burg B, Siegler MA, Saez Talens V, Kieltyka RE, Bonnet S. Turning on in Four Soldenella Species. Phytochemical Anal. 28, 567-574 (2017). the red phosphorescence of a [Ru(tpy)(bpy)(Cl)]Cl complex by amide Hartmann A, Murauer A, Ganzera M. Quantitative analysis of mycospo- substitution: self-aggregation, toxicity, and cellular localization of an rine-like amino acids in marine algae by capillary electrophoresis with emissive ruthenium-based amphiphile. Chemical Commun. 53, 11126- diode-array detection. J. Pharm. Biomed. Anal. 138, 153-157 (2017). 11129 (2017). Hochleitner J, Akram M, Ueberall M, Davis RA, Waltenberger B, Skhirtladze A, Nebieridze V, Benidze M, Kemertelidze E, Ganzera M. Stuppner H, Sturm S, Ueberall F, Gostner JM, Schuster D. A combinato- Furostanol glycosides from the roots of Tribulus terrestris L. Bulletin of rial approach for the discovery of cytochrome P450 2D6 inhibitors from the Georgian National Academy of Sciences 11, 122-126 (2017). nature. Sci. Rep. 7, 8071 (2017). Skhirtladze AV, Kopaliani TA, Nebieridze VG; Kemertelidze EP, Ganzera Kemertelidze E, Skhirthladze M, Ganzera M. Furostanol and triterpene M. New steroidal glycosides from pericarp of Digitalis ferruginea. saponins from the roots of Digitalis ciliata. Chemistry of Natural Com- Chemistry of Natural Compounds 53, 1083-1087 (2017). pounds 53, 492-496 (2017). Taibon J, Sturm S, Strasser H, Stuppner H. Combination of a QuEChERS- Khan SY, Awad EM, Oszwald A, Mayr M, Yin X, Waltenberger B, based extraction protocol with a fast and selective UHPLC-QTOF-MS Stuppner H, Lipovac M, Pavel U, Breuss JM. Premature senescence of assay for the detection and quantification of Metarhizium brunneum endothelial cells upon chronic exposure to TNFα can be prevented by metabolites from honey samples. SOJ Pharmacy & Pharmaceutical Sci- N-acetyl cysteine and plumericin. Sci. Rep. 7, 39501 (2017). ences 4, 1-5 (2017). 88  Publications Publications  89

Papadakis ES, Robson N, Yeomans A, Bailey S, Laversin S, Beer S, Sayan A, Ashton-Key M, Schwaiger S, Stuppner H, Troppmair J, Packham G, Cutress R. A combination of trastuzumab and BAG-1 inhibition synergistically targets HER2 positive breast cancer cells. Oncotarget 7, Temml V, Garscha U, Romp E, Schubert G, Gerstmeier J, Kutil Z, Ma- 18851-18864 (2016). tuszczak B, Waltenberger B, Stuppner H, Werz O, Schuster D. Discovery Papadakis ES, Barker CR, Syed H, Reeves T, Schwaiger S, Stuppner H, of the first dual inhibitor of the 5-lipoxygenase-activating protein and Troppmair J, Blaydes JP, Cutress RI. The Bag-1 inhibitor, Thio-2, reverses soluble epoxide hydrolase using pharmacophore-based virtual screen- an atypical 3D morphology driven by Bag-1L overexpression in a MCF- ing. Sci. Rep. 7, 42751 (2017). 10A model of ductal carcinoma in situ. Oncogenesis 5, e215 (2016). Vrabl P, Schinagl CW, Artmann DJ, Krüger A, Ganzera M, Pötsch A, Pfeifer I, Murauer A, Ganzera M. Determination of coumarins in the Burgstaller W. The Dynamics of Plasma Membrane, Metabolism and roots of Angelica dahurica by supercritical fluid chromatography. J. Respiration (PM-M-R) in Penicillium ochrochloron CBS 123824 in Pharm. Biomed. Anal. 129, 246-251 (2016). Response to Different Nutrient Limitations—A Multi-level Approach Schäfer S, Salcher S, Seiter M, Ranninger C, Moest M, Obexer P, Huber to Study Organic Acid Excretion in Filamentous Fungi. Frontiers in CG, Ausserlechner MJ, Schwaiger S, Stuppner H. Characterization of Microbiol. 8, 2475 (2017). the XIAP-Inhibiting Proanthocyanidin Fraction of the Aerial Parts of Vuorinen A, Engeli RT, Leugger S, Bachmann F, Akram M, Atanasov AG, Ephedra sinica. Planta Med. 82, 973-985 (2016). Waltenberger B, Temml V, Stuppner H, Krenn L, Ateba SB, Njamen D, Schaible AM, Filosa R, Krauth V, Temml V, Pace S, Garscha U, Liening S, Davis RA, Odermatt A, Schuster D. Potential Antiosteoporotic Natural Weinigel C, Rummler S, Schieferdecker S, Nett M, Peduto A, Collarile Product Lead Compounds That Inhibit 17β-Hydroxysteroid Dehydroge- S, Scuotto M, Roviezzo F, Spaziano G, De Rosa M, Stuppner H, Schuster nase Type 2. J. Nat. Prod. 80, 965-974 (2017). D, D´Agostino B, Werz O. The 5-lipoxygenase inhibitor RF-22c potently Alsabil K, Suor-Cherer S, Koeberle A, Viault G, Lavaud A, Temml V, suppresses leukotriene biosynthesis in cellulo and blocks bronchocon- Waltenberger B, Schuster D, Litaudon M, Lorkowski S, de Vaumas R, striction and inflammation in vivo. Biochem. Pharmacol. 112, 60-71

publications | Helesbeux JJ, Guilet D, Stuppner H, Werz O, Seraphin D, Richomme P. (2016). Semisynthetic and Natural Garcinoic Acid Isoforms as New mPGES-1 Scharinger B, Messner B, Türkcan A, Schuster D, Vuorinen A, Pitterl F, Inhibitors. Planta Med. 82, 1110-1116 (2016). Heinz K, Arnhard K, Laufer G, Grimm M, Stuppner H, Oberacher H, Antal DS, Pinzaru I, Borcan F, Marti TD, Ledeti I, Coricovac D, Schwaiger Eller P, Ritsch A, Bernhard D. Leoligin, the major lignan from edelweiss, S, Stuppner H, Dehelean CA, Ollivier E, Soica C. Inclusion Complexes of inhibits 3-hydroxy-3-methyl-glutaryl-CoA reductase and reduces choles- the Aurone Sulfuretin and the Chalcone Butein from Cotinus coggygria terol levels in ApoE −/− mice. J. Mol. Cell. Cardiol. 99, 35-46 (2016). Wood in Two Cyclodextrin Types: First data on physico-chemical proper- Skhirtladze A, Kemertelidze E, Nebieridze V, Ganzera M. Phenyletha- ties. Revista De Chimie 67, 1104-1109 (2016). noid Glycosides from the Roots of Digitalis ciliata Trautv. Helvetica Becker K, Hartmann A, Ganzera M, Fuchs D, Gostner JM. Immuno- Chimica Acta 99, 241-245 (2016). modulatory Effects of the Mycosporine-Like Amino Acids Shinorine and Waltenberger B, Garscha U, Temml V, Liers J, Werz O, Schuster D, Porphyra-334. Mar. Drugs 14, 119 (2016). Stuppner H. Discovery of Potent Soluble Epoxide Hydrolase (sEH) Inhibi- Cheilari A, Sturm S, Intelmann D, Seger C, Stuppner H. Head-to-Head tors by Pharmacophore-Based Virtual Screening. Journal of Chemical Comparison of Ultra-High-Performance Liquid Chromatography with Information and Modeling 56, 747-762 (2016). Diode Array Detection versus Quantitative Nuclear Magnetic Resonance Wang L, Ladurner A, Latkolik S, Schwaiger S, Linder T, Hosek J, Palme V, for the Quantitative Analysis of the Silymarin Complex in Silybum mari- Schilcher N, Polanský O, Heiss EH, Stangl H, Mihovilovic MD, Stuppner anum Fruit Extracts. J. Agric. Food Chem. 64, 1618-1626 (2016). H, Dirsch VM, Atanasov AG. Leoligin, the Major Lignan from Edelweiss Hartmann A, Holzinger A, Ganzera M, Karsten U. Prasiolin, a new (Leontopodium nivale subsp. alpinum), Promotes Cholesterol Efflux UV-sunscreen compound in the terrestrial green macroalga Prasiola from THP-1 Macrophages. J. Nat. Prod. 79, 1651-1657 (2016). calophylla (Carmichael ex Greville) Kützing (Trebouxiophyceae, Chlo- Winderl B, Schwaiger S, Ganzera M. Fast and improved separation of rophyta). Planta 243, 161-169 (2016). major coumarins in Ammi visnaga (L.) Lam. by Supercritical Fluid Chro- Heiss EH, Liu R, Waltenberger B, Khan S, Schachner D, Kollmann P, Zim- matography. J. Sep. Sci. 39, 4042-4048 (2016). mermann K, Cabaravdic M, Uhrin P, Stuppner H, Breuss JM, Atanasov Zatelli GA, Temml V, Kutil Z, Landa P, Vanke T, Schuster D, Falkenberg AG, Dirsch VM. Plumericin inhibits proliferation of vascular smooth M. Miconidin Acetate and Primin as Potent 5-Lipoxygenase Inhibitors muscle cells by blocking STAT3 signaling via S-glutathionylation. Sci. from Brazilian Eugenia hiemalis (Myrtaceae). Planta Medica Letters 3, Rep. 6, 20771 (2016). e17-e19 (2016). 90  Publications Publications  91

Ahammer L, Grutsch S, Kamenik AS, Liedl KR, Tollinger M. Structure of the major apple allergen Mal d 1. J. Agric. Food Chem. 65, 1606-1612 (2017). M. Tollinger, C. Kreutz Nussbaumer F, Juen M, Gasser C, Kremser J, Mueller T, Tollinger M, Juen MA, Wunderlich CH, Nussbaumer F, Tollinger M, Kontaxis G, Kon- Kreutz CK. Synthesis and incorporation of 13C-labeled DNA building rat R, Hansen DF, Kreutz CK. Excited states of nucleic acids probed by blocks to probe structural dynamics of DNA by NMR. Nucleic Acids Res. proton relaxation dispersion NMR Spectroscopy. Angew. Chem. Int. Ed. 45, 9178-9192 (2017). Engl. 55, 12008-12012 (2016). Kremser J, Strebitzer E, Plangger R, Juen MA, Nußbaumer F, Glasner H, Moschen T, Grutsch S, Juen, MA, Wunderlich CH, Kreutz C, Tollinger M. Breuker K, Kreutz C. Chemical synthesis and NMR spectroscopy of long Measurement of ligand-target residence times by 1H relaxation disper- stable isotope labelled RNA. Chem. Commun. 53, 12938-12941 (2017). sion NMR spectroscopy. J. Med. Chem. 59, 10788-10793 (2016). Rennella E, Sara T, Juen M, Wunderlich C, Imbert L, Solyom Z, Favier A, Zhou H, Kimsey IJ, Nikolova EN, Sathyamoorthy B, Grazioli G, McSally Ayala I, Weinhaeupl K, Schanda P, Konrat R, Kreutz C, Brutscher B. RNA J, Bai T, Wunderlich CH, Kreutz C, Andricioaei I, Al-Hashimi HM. M(1)A binding and chaperone activity of E. coli cold-shock protein CspA. Nucl. and m(1)G disrupt A-RNA structure through the intrinsic instability of Acids Res. 45, 4255-4268 (2017). Hoogsteen base pairs. Nat. Struct. Mol. Biol. 23, 803-810 (2016). Grutsch S, Fuchs JE, Ahammer L, Kamenik AS, Liedl KR, Tollinger M. Grutsch S, Brüschweiler S, Tollinger M. NMR methods to study dynamic Conformational flexibility differentiates naturally occurring Bet v 1 allostery. PLoS Comput. Biol. 12, e1004620 (2016). isoforms. Int. J. Mol. Sci. 18, 1192 (2017). Machado Y, Freier R, Scheiblhofer S, Thalhamer T, Mayr M, Briza P, Ahammer L, Grutsch S, Wallner M, Ferreira F, Tollinger M. NMR reso- Grutsch S, Ahammer L, Fuchs J, Wallnöfer H, Isakovic A, Kohlbauer V, nance assignments of a hypoallergenic isoform of the major birch pol- Hinterholzer A, Steiner M, Danzer M, Horejs-Hoeck J, Ferreira F, Liedl len allergen Bet v 1. Biomol. NMR Assign. 11, 231-234 (2017). KR, Tollinger M, Lackner P, Johnson CM, Brandstetter H, Thalhamer Neuner S, Kreutz C, Micura R. The synthesis of 15N7-Hoogsteen face-

publications | J, Weiss R. Fold-stability during endolysosomal acidification is a key labeled adenosine amidite for solid phase RNA synthesis. Monatsh. factor for allergenicity and immunogenicity of the major birch pollen Chem. 148, 149-155 (2017). allergen. J. Allergy Clin. Immunol. 137, 1525-1534 (2016). Wolter AC, Weickhmann AK, Nasiri AH, Hantke K, Ohlenschläger O, Duchardt-Ferner E, Juen M, Kreutz C, Wöhnert J. NMR resonance as- Wunderlich CH, Kreutz C, Duchardt-Ferner E, Wöhnert J. A Stably Pro- signments for the tetramethylrhodamine binding RNA aptamer 3 in tonated Adenine Nucleotide with a Highly Shifted pKa Value Stabilizes complex with the ligand 5-carboxy-tetramethylrhodamine. Biomol. the Tertiary Structure of a GTP-Binding RNA Aptamer. Angew. Chem. NMR Assign. 11, 29-34 (2016). Int. Ed. Engl. 56, 401-404 (2017). Ahammer L, Grutsch S, Tollinger M. NMR resonance assignments of the Tants JN, Fesser S, Kern T, Stehle R, Geerlof A, Wunderlich CH, Juen MA, major apple allergen Mal d 1. Biomol. NMR Assign. 10, 287-290 (2016). Hartlmüller C, Böttcher R, Kunzelmann S, Lange O, Kreutz C, Förste- Ren A, Vusurovic N, Gebetsberger J, Gao P, Juen M, Kreutz C, Micura mann K, Sattler M. Molecular basis for asymmetry sensing of siRNAs by R, Patel DJ. Pistol ribozyme adopts a pseudoknot fold facilitating site- the Drosophila Loqs-PD/Dcr-2 complex in RNA interference. Nucl. Acids specific in-line cleavage. Nat. Chem. Biol. 12, 702-708 (2016). Res. 45, 12536-12550 (2017). Scherl M, Müller T, Kreutz C, Huber RG, Zass E, Liedl KR, Kräutler B. Vuorinen A, Engeli RT, Leugger S, Kreutz C, Schuster D, Odermatt Chlorophyll catabolites in fall leaves of the wych elm tree present a A, Matuszczak B. Phenylbenzenesulfonates and -sulfonamides as novel glycosylation motif. Chemistry 22, 9498-9503 (2016). 17β-hydroxysteroid dehydrogenase type 2 inhibitors: Synthesis and Longhini AP, LeBlanc RM, Becette O, Salguero C, Wunderlich CH, John- SAR-analysis. Bioorg. Med. Chem. Lett. 27, 2982-2985 (2017). son BA, D’Souza VM, Kreutz C, Dayie TK. Chemo-enzymatic synthesis of Führer S, Ahammer L, Ausserbichler A, Scheffzek K, Dunzendorfer-Matt site-specific isotopically labeled nucleotides for use in NMR resonance T, Tollinger M. NMR resonance assignments of the EVH1 domain of assignment, dynamics and structural characterizations. Nucleic Acids Neurofibromin’s recruitment factor Spred1. Biomol. NMR Assign. 11, Res. 44, e52 (2016). 305-308 (2017). 92  CMBI news CMBI news  93

CMBI - careers of young researchers

Daniela Schuster was awarded one of the first two Ingeborg Hochmair Jerome Mertens joined the Institute of Molecular | Professorships by the University of Innsbruck. This temporary professor- Biology in 2017 starting his tenure track, and is ship is awarded to highly successful, young female scientists to increase currently building his research group at the De- the number of female professors in faculties, where female scientists partment for Genomics, Stem Cell Biology and are underrepresented. This position should also help the awardees to be Regenerative Medicine. Dr. Mertens is also con- appointed full professors at other institutions. In 2018, Daniela Schuster ducting his research as a staff scientist at the Salk follows a call as Professor for Pharmaceutical and Medicinal Chemistry at Institute for Biological Studies in La Jolla, CA, USA. the Paracelsus Medical Private University of Salzburg. Together with her His work combines human neural cell reprogram- team, she will build up teaching courses and infrastructure for the newly ming technologies such as the direct neuronal conversion (iN) with next- established pharmacy study and continue her research on in silico acti- generation sequencing strategies, bioinformatics, neuronal cell biology vity profiling, especially in the field of environmental chemical toxicity. and functional neuroscience. With a focus on age-related neurodege- nerative diseases and neuropsychiatric disorders, his goal is to unravel the impact of epigenetic cellular states on neuronal pathology, and to direct the interphase between aging and disease. In 2017, he has been awarded the prestigious K99 Pathway to Independence award from the National Institute for Aging. faculty appointment 94  CMBI news CMBI news  95

CMBI - careers of young researchers

In 2013, the CMBI initiated an annual call for a young investigator David Granig (Simone B. Sartori and Nicolas | scholarship. Aim of this scholarship is to support a young career Singewald Group, Institute of Pharmacy, (Master/PhD level) and to strengthen collaboration within the CMBI Department of Pharmacology and Toxicology): by selecting a candidate, who is engaged in a high-profile research Discovery and characterization of non- project connecting two or more CMBI member groups. The scholar and peptidergic neuropeptide S receptor agonists as his project are supported by allocating prize money for expendable potential novel anxiolytics materials and consumables. In collaboration with the Dept. of Pharmacognosy Previous CMBI Scholars: (Prof. Stuppner, Dr. Schwaiger) four derivatives of our previously 2013: R. Spitzer identified potential neuropeptide S receptor (NPSR) agonist were (Christoph Kreutz Group, Department of Organic Chemistry) synthesized in house, virtually screened (Prof. Daniela Schuster, 2014: Johanna E. Mayrhofer Dept. Pharmaceutical Chemistry) and characterized in-vitro using (Eduard Stefan Group, Department of Biochemistry) a refined cell-based Ca2+-mobilization assay. The parent compound 2015: Julian Wimmer (Ilse Kranner, Department of Botany) and its two dimethyl- and diethyl-derivatives demonstrate specific 2016: Florian Enzler (Eduard Stefan, Department of Biochemistry) agonistic properties at the NPSR with moderate efficacy. Since these two derivatives are suggested to display BBB penetrance, they represent leads for the development of small NPSR agonists which are The current CMBI Scholars for 2017: expected to display similar unique characteristics as the parent NPS, namely to induce anxiolytic effects without any signs of sedation. The

Anna Hausruckinger (Frank Edenhofer Group, collaborations are ongoing in terms of refining the pharmacophore Department of Molecular Biology): CaV expression model, the synthesis of derivatives and in vivo testing for potential in neurons derived from human iPS cells therapeutic effects.

Voltage-gated calcium channels (CaVs) are involved in the regulation of various neuronal functions in the central nervous system, such as CMBI scholarships neurotransmitter secretion, postsynaptic signal integration and neuronal plasticity. Current research mainly depends on the analyses of heterologous expression systems (such as HEK cells) or primary animal neurons that are far from being ideal to correlate with the physiology of human neurons. Induced pluripotent stem (iPS) cells represent a promising alternative source of human neurons. The overall aim of the project is to elaborate on an expression analysis of CaVs subunits in neurons derived from human iPS cells. Moreover, we investigate to which extent the modulation of CaV functionality by subunit overexpression will impact on the maturation and synaptogenesis of in vitro differentiated neurons. 96  Awards and honors for CMBI scientists Awards and honors for CMBI scientists  97

The successful scientific activities of the CMBI are reflected in numerous Katharina Günther, Department of Molecular Biology: awards and honors received from several members during the reporting Poster Prize, 9th ÖGMBT Annual Meeting & 8th Life Science Meeting, period. "Molecular and cellular mechanisms of human diseases". Innsbruck, 2017 Nadja Hofer, Department of Pharmacology and Toxicology: Würdigungspreis Bundesministerium für Wissenschaft, Forschung und Wirtschaft (2016) Ilse Kranner, Department of Botany President of the ATSPB (Austrian Society of Plant Biology) and

Katharina Günther national delegate on the FESPB (Federation of European Societies of Plant Biology) Council Johannes Kremser, Department of Organic Chemistry: "Suraj Manrao Poster Award" at the 20th ISMAR in Quebec City, awards | Canada, July 2017. Birgit Lengerer, Department of Zoology Best Poster Award (3rd Place), Life Science PhD Meeting Innsbruck 2017 Awards and honors for CMBI scientists Carina Miggitsch, Research Department for Biomedical Ilse Kranner Linda Ahammer, Department of Organic Chemistry: Aging Research: "Young Investigator Award" at the 27th ICMRBS in Kyoto, Japan, Best Short Talk Award, Life Science PhD Meeting Innsbruck 2017 August 2016. Simon Moosmang, Department of Pharmacognosy: Erwann Arc, Department of Botany: Young Talent Poster Award, First Place - Best Poster, VASCage Best lecture of an early career scientist, 21th meeting of the ATSPB, Meeting, 2017 Erwann Arc Berchtesgaden, 2016 Sandro Neuner, Department of Organic Chemistry Giorgia Baraldo, Research Department for Biomedical Aging Best Paper Award 2017, Monatshefte für Chemie/Chemical Monthly, Research: Springer Grant D. Swarovski AG, “Plant-derived compounds for inhibition of Michael Neustetter, Department of Ion Physics and Applied Physics: NOX4 to counteract vascular aging” Award of Excellence of the Austrian Federal Ministry of Science and Giovanni Calderisi, Department of Organic Chemistry: Research (2017). Student Travel Award of the American Society for Mass Nadine Ortner, Department of Pharmacology and Toxicology: Spectrometry for presentation of a poster at the 65th ASMS Principality of Liechtenstein Prize (2017) conference, Indianapolis, IN, USA (2017) Nadine Ortner, Department of Pharmacology and Toxicology: Sebastian Führer, 17. Österreichische Chemietage, Univ. Salzburg (2017), Foto: Göch Andreas Friedl, Department of Pharmacognosy: Jubiläumsfonds der Universität Innsbruck und der Medizinischen Meda Preis für Phytopharmaka-Forschung 2016 Universität Innsbruck zur Förderung wissenschaftlicher Sebastian Führer, Department of Organic Chemistry: Kooperationsprojekte (2017) GÖCH-Förderungspreis für die besten Diplomarbeiten (Gesellschaft Nadine Ortner, Department of Pharmacology and Toxicology: Österreichischer Chemiker), Austria, 2017 Innsbruck 2017. ALUMNI-I-MED Talk Prize, Life Science PhD Meeting (2017) Marco Grasse, Research Department for Biomedical Aging Research: Luca Pangrazzi, Research Department for Biomedical Grant D. Swarovski AG 2017, “Effects of GM-CSF application Aging Research: during tetanus/diphtheria vaccination on the antigen-specific T cell Grant D. Swarovski AG 2016, “Characterization of niche providing response in vivo” Marco Grasse Luca Pangrazzi cells in the bone marrow” 98  Awards and honors for CMBI scientists CMBI - meetings and seminar series  99

CMBI - meetings

LIFE SCIENCE PHD MEETING This meeting was held in the Center for Chemistry and Biomedicine in Isabella Pfeifer, Department of Pharmacognosy: Innsbruck on April 18-19, 2017. It brings together the annual meetings Meda Preis für Phytopharmaka-Forschung 2017 of the different FWF-funded excellence doctoral programs and, most Michael Schauperl, Department of General, Inorganic and importantly is organized by PhD students. The participating programs Theoretical Chemistry: were MCBO, SPIN, and HOROS but all PhD students of both universities Dr. Otto Seibert Science Award, 11/2017 were invited. The student organizers also brought in excellent plenary Michael Schauperl, Department of General, Inorganic and speakers: Catherina Becker (University of Edinburgh, UK), Jörg Köhl Theoretical Chemistry: (University of Lübeck, DE) and Arnoud Sonnenberg (The Netherlands Science Award 2017 of the Tyrolean Economic Chamber, 11/2017 Cancer Institute, Amsterdam, NL). Of course, CMBI many CMBI PhD Daniela Schuster, Department of Pharmaceutical Chemistry: students and postdocs alo participated and ppresented their recent re- Ingeborg Hochmair Professorship 2016 search findings. Some of them won prizes, such as Nadine Ornter (Phar- Michael Schauperl Daniela Schuster, Department of Pharmaceutical Chemistry: macology, Pharmacy) for an excellent oral presentation (together with Poster award (2nd rank) at Pharma 2030 jury member Prof. Raimund Margreiter). Torsten Schwerte, Department of Zoology:

Edmund Optics Educational Award Winner in Europe (2017) Anita Siller, Department of Pharmacology and Toxicology: Studienförderpreis des Deutschen Freundeskreises der Universitäten in Innsbruck e. V. (2016) Anita Siller, Department of Pharmacology and Toxicology: Best Poster Award, 9th ÖGMBT Annual Meeting (2017) Anita Siller, Department of Pharmacology and Toxicology: Best Poster Award, 8th Life Science Meeting (2a017) Anita Siller, Department of Pharmacology and Toxicology:

Würdigungspreis des Bundesministeriums für Wissenschaft, annual meeting | Wirtschaft und Forschung (2017) Hermann Stuppner, Department of Pharmacognosy: Wissenschaftspreis für außergewöhnliche Forschungsleistung der Stiftung Südtiroler Sparkasse, 2017 Hermann Stuppner, Department of Pharmacognosy: Bruker Award, 2017 Hermann Stuppner Birgit Waldner, Department of General, Inorganic and

Theoretical Chemistry: 23RD SCIENTIFIC SYMPOSIUM Poster award of the Molecular Modeling Workshop Erlangen OF THE AUSTRIAN PHARMACOLOGICAL SOCIETY APHAR Alexander Weiss, Research Department for Biomedical Aging CMBI members are also part of the Austrian Pharmacological Society Research: APHAR and were in charge of organizing the annual APHAR meeting in Grant D. Swarovski AG, “Struktur- und Aktivitätsbestimmung des Innsbruck on September 28–29, 2017. In addition to regulatory aspects menschlichen Stoffwechselenzyms FAHD2a” of the European Medicines Agency (plenary lecture by Thomas Salmon- Julia Wunderer, Department of Zoology sen, London & Medicinal Products Agency, Uppsala, SWE) John A. Ci- Best Poster Award (1st Place), Life Science PhD Meeting Innsbruck 2017 dlowski (Natl Inst. of Environmental Health Services, USA) and Michel Julia Wunderer, Department of Zoology Bouvier (Univ. de Montréal, CAN) also presented recent work not only Best Short Talk Award (1st Place), 6th Annual CMBI Meeting 2016 Hermann Stuppner interesting for pharmacologists but also for the CMBI community. 100  CMBI - meetings and seminar series CMBI - meetings and seminar series  101

6TH ANNUAL CMBI MEETING INNSBRUCK 2016 On March 3rd and 4th, the CMBI held its 6th annual joint meeting in Gnadenwald focusing on drugs and biotechnology, proteomics and me- tabolomics, protein structure, signaling, disease and stress mechanisms. About 120 – mostly young – researchers from CMBI member labs gath- ered in order to share their newest Life Science findings in more than 70 oral and poster presentations and to initiate discussion and interaction. Three impressive, high-profile plenary speakers, Christian Griesinger (MPI Göttingen), Remco Sprangers (MPI Tübingen), Almut Schulze (Uni Würzburg), and an inspiring Introduction Lecture held by the newly ap- pointed faculty member Frank Edenhofer complemented the program. As in previous years, three prizes were selected with the help of the plenary speakers for the best young scientists’ conference presenta- tions. Two poster prizes were awarded to Marina Frener (Department for Organic Chemistry) and Armin Wilfinger (Department of Molecular 9TH ÖGMBT ANNUAL MEETING 2017 & Biology). The award for the best short talk was given to Julia Wunderer 8TH LIFE SCIENCE MEETING INNSBRUCK (Department of Zoology). This largest regular life science meeting in western Austria took place in Innsbruck from September 25–27, 2017. It also serves as a key annual meeting of the biomedical and biological research community of the two Innsbruck universities. The CMBI groups also actively participated with interesting oral and poster presentations thereby increasing the critical mass of the event. All enjoyed the interdisciplainary conversa- tions with scientists from all over Austria. Excellent invited speakers also particpated with the lively discusisons at the posters. Invited speakers were Andrea Ballabio, TIGEM, IT; Thomas Carell, LMU, DE; Christine Falk, MHH, DE; Micheala Frye, University of Cambridge, UK; Florian Greten, FCI, DE; Silvio Gutkind, UCSD, US; Karl Lohner, KFU Graz, AT; Alexander Mankin, UIC, US; Richard Marais, University of Manchester, UK; Alexan- der Meissner, Harvard University, US; Maria M. Mota, iMM Lisboa, PT; Matthias Peter, ETH Zürich, CH; Steven Taylor, University of Manchester, UK; Andreas Trumpp, DKFZ, DE; Georg Winter, CeMM, AT; Nieng Yan, Tsinghua University, CN. Students and young scientists from several FWF- funded excellence consortia participated at this meeting, including the doctoral programs SPPIN, HOROS and MCBO as well as the neuroscience research consortium SFB-F44. CMBI investigators actively participate in all these programs. annual meeting | 102  CMBI - meetings and seminar series CMBI - meetings and seminar series  103

CMBI - meetings

5th Life Science Meeting Innsbruck, Innsbruck, Tyrol, Sept. 25th - 27th, 2013 >> Asifa Akhtar Max Planck Institute of Immunobiology and Epigenetics, Germany >> Michel Desjardins, Universite de Montreal, Canada >> Carl-Philipp Heisenberg, IST Austria >> Karolin Luger, Colorado State University, USA >> Frauke Melchior, ZMBH Heidelberg University, Germany >> Nikolaus Pfanner, University of Freiburg, Germany >> Britta Qualmann, Friedrich-Schiller-University Jena, Germany >> Elena Rugarli, University of Cologne, Germany >> Susan S. Taylor, University of California San Diego, USA

4th Life Science Meeting Innsbruck, Igls, Tyrol, Sept. 27th - 28th, 2012 >> Christine Foyer External speakers at previous CMBI meetings Centre for Plant Sciences, University of Leeds, United Kingdom >> Ari Helenius, Institute of Biochemistry, ETH Zürich, Switzerland In addition to the many exciting short talks and poster presentations from >> Anne-Claude Gavin members of the CMBI and the Biocenter, guest lectures from invited top Structural and Computational Biology, EMBL Heidelberg, Germany scientists contributed to the lively discussions. Like in previous meetings, the speakers also served as advisory experts for our research activities and 3rd Life Science Meeting Innsbruck, Igls, Tyrol, Sept. 23th - 24th, 2011 as referees for the poster awards for young scientists. >> Ilme Schlichting Department of Biomolecular Mechanisms, Max Planck Institute for Medical 6th CMBI Meeting Innsbruck, Innsbruck, Tyrol, March 3th - 4th, 2016 Research, Heidelberg, Germany >> Christian Griesinger >> Adrian R. Ferré-D’Amaré Max Planck Institute for Biophysical Chemistry Göttingen, Germany Laboratory of RNA Biophysics and Cellular Physiology, Biochemistry and >> Frank Edenhofer, University of Innsbruck, Austria Biophysics Center, National Heart, Lung and Blood Institute, Bethesda, USA >> Remco Sprangers, Max Planck Campus Tübingen, Germany >> Daniel Minor >> Almut Schulze, University of Wuerzburg, Germany Cardiovascular Research Institute, Departments of Biochemistry & Biophysics, and Cellular & Molecular Pharmacology California Institute for Quantitative 7th Life Science Meeting Innsbruck, Innsbruck, Tyrol, Feb. 27th, 2015 Biomedical Research, University of California, San Francisco, USA >> Peter Hinterdorfer, University of Linz, Austria >> Eduard Stefan, University of Innsbruck, Austria 2nd Life Science Meeting Innsbruck, Igls, Tyrol, Sept. 24th - 25th, 2010 >> Peter Ladurner, University of Innsbruck, Austria >> Maria Sibilia >> Gunter Meister, University of Regensburg, Germany Institute for Cancer Research, Medical University of Vienna, Austria >> Gerald Obermair, Medical University of Innsbruck, Austria >> Adriano Aguzzi >> Natascha Kleiter, Medical University of Innsbruck, Austria Institute of Neuropathology, University Hospital of Zürich, Switzerland >> Pascal Meier, ICR London, UK >> Rik Korswagen

annual meeting | >> Marlies Meisl, University of Chicago, USA Hubrecht Institute, Utrecht, Netherlands >> Martin Puhr, Medical University of Innsbruck, Austria >> Bill Earnshaw, University of Edinburgh, UK 1st Life Science Meeting Innsbruck, Igls, Tyrol, Sept. 18th - 19th, 2009 >> Dirk Trauner 6th Life Science Meeting Innsbruck, Innsbruck, Tyrol, Sept. 24th - 25th, 2014 Department of Chemistry and Biochemistry, LMU Munich, Germany >> Robert T. Batey, University of Colorado Boulder, USA >> Didier Stainier, University of California, San Francisco, USA >> Veronika Sexl, University of Veterinary Medicine, Vienna, Austria >> Wolfgang Baumeister >> Florian Kronenberg, Medical University of Innsbruck, Austria Max-Planck-Institute of Biochemistry, Martinsried, Germany 104  CMBI - meetings and seminar series CMBI - meetings and seminar series  105

CMBI - Meetings and Seminar Series CMBI - meetings CMBI - seminar series

The CMBI seminars are a very important integrative and multidisciplinary ac- tivity of the CMBI and are also part of the PhD programs established within the CMBI. So far, it hosted 111 lectures from renowned scientists from the US, Canada, Australia, Sweden, Denmark, France, Italy, UK, Ireland, Netherlands, Germany, Belgium, Switzerland, and Austria. It also provides a forum for ex-

5th Annual CMBI-Meeting Igls, Tyrol, Sept. 26th - 27th, 2008 cellent scientists from the Innsbruck Universities. Seminar speakers of the last >> Peter Hegemann three years are listed here: Institute for Biology, Experimental Biophysics, Humboldt University Berlin, Germany 2017 >> Stefan Schulte-Merker Rasmus Linser, Ph.D. Hubrecht Laboratory, Netherlands Institute for Developmental Biology Department for Chemistry, Ludwig-Maximilians-University of Munich, Utrecht, Netherlands Germany Michel Bouvier, Ph.D.

4th Annual CMBI-Meeting Igls, Tyrol, Sept. 28th - 29th, 2007 F.C.A.H.S., FRSC Université de Montréal, Institute of Research in seminars | >> Ulf R. Rapp Immunology and Cancer, Montréal, Canada Medical Radiation & Cell Research, Univ. of Würzburg, Germany Oliver Rocks, Ph.D. >> Gregory J. Kaczorowski Max Delbrück Center for Molecular Medicine, Berlin, Germany Merck Research Laboratories, Rahway, New Jersey, USA David Gillespie, Ph.D. >> Thomas W. Holstein Institute of Biomedical Technologies, Centre for Biomedical Research Institute of Zoology, University of Heidelberg, Germany of the Canary Islands Nathan Lüdtke, Ph.D.

3rd Annual CMBI-Meeting Vill, Tyrol, Sept. 29th - 30th, 2006 University of Zurich, Department of Chemistry, Zurich, Switzerland >> Naweed I. Syed, Anatomy and Physiology, University of Calgary, Canada David Henshall, Ph.D. >> Erwin F. Wagner Professor of Molecular Physiology and Neuroscience, Royal College of Research Institute of Molecular Pathology, Vienna, Austria Surgeons in Ireland, Dublin, Ireland >> Walter Schaffner Helmut Schwarz, Ph.D. Institute of Molecular Biology, University of Zurich, Switzerland Institute of Chemistry, Technical University of Berlin, Germany Hans Schöler, Ph.D.

2nd Annual CMBI-Meeting Vill, Tyrol, Sept. 30th - Oct. 1st, 2005 Max-Planck Institute for Molecular Biomedicine, Münster, Germany annual meeting | >> Wolfram Saenger Andrea Pauli, Ph.D. Inst. of Chemistry & Crystallography, Free University Berlin, D IMP - Research Institute of Molecular Pathology, Vienna, Austria >> Peter Herrlich, Institute of Molecular Biotechnology, Jena, Germany Monika Hassel, Ph.D. >> Elisabeth Knust, Institute of Genetics, University of Düsseldorf, Germany Morphology and Evolution of Invertebrates, Philipps-University of Marburg, Germany

1st Annual CMBI-Meeting Vill, Tyrol, Oct. 1st - 2nd, 2004 >> Robert Huber 2016 Nobel Laureate in Chemistry, MPI of Biochemistry, Martinsried, D Stephan A. Sieber, Ph.D. >> Reinhard Fässler Department of Chemistry, Technical University of Munich, Germany Max-Planck-Institute of Biochemistry, Martinsried, Germany Dirk Trauner, Ph.D. >> Daniela Pietrobon Department for Chemistry and Biochemistry, Ludwig-Maximilians- Dept. of Biomedical Sciences, University of Padova, Italy University of Munich, Germany 106  CMBI - meetings and seminar series

CMBI - seminar series

Jeffrey Liu, Ph.D. Max Planck Institute of Biochemistry, Martinsried, Germany Alois Fürstner , Ph.D. Organometallic Chemistry, Max-Planck-Institute, Mühlheim, Germany Dominique Massotte, Ph.D. Institut des Neurosciences Cellulaires et Intégratives, Strasbourg, France Michael Heise, Ph.D. South Westphalia University of Applied Sciences, Iserlohn, Germany Norbert Bischofberger, Ph.D. Gilead Sciences, California, USA Gerald Schwank, Ph.D. Institute for Molecular Health Sciences, ETH Zurich, Switzerland Rudi Vennekens, Ph.D. seminars | Laboratory of Ion Channel Research Biology, KU Leuven, NL Frank Edenhofer, Ph.D. Institute of Molecular Biology, University of Innsbruck, A Jutta Schüller, Ph.D. Taconic Biosciences, Cologne, G Roland Sigel, Ph.D. Department of Chemistry, University of Zurich, CH

2015 Stefan Hörtensteiner, Ph.D. Institute of Plant Biology, University of Zurich, CH Georg S. Baillie, Ph.D. Molecular Pharmacology, Institute of Cardiovascular and Medical Sciences, University of Glasgow, UK Daniel Gerlich, Ph.D. IMP, Vienna, AT Elena Conti, Ph.D. Director, MPI, Munich, Germany Alexander Stark, Ph.D. IMP, Vienna, AT Markus Ralser, Ph.D. CSBC, University of Cambridge, UK Matthias Hentze, MD EMBL, Heidelberg, Germany Roger Schibli, Ph.D. Institute for Pharmaceutical Sciences, ETH Zurich, Switzerland contact |

Centrum für Molekulare Biowissenschaften Innsbruck Universität Innsbruck

Office: Innrain 80-82a A-6020 Innsbruck Tel: +43 512 507-57501 Fax: +43 512 507-57599 [email protected] www.uibk.ac.at/cmbi