DOCSLIB.ORG

Analysis of the Novel Lipid Transfer Protein Anchored at Membrane Contact Sites (LAM) Family

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Analysis of the Novel Lipid Transfer Protein Anchored at Membrane Contact Sites (LAM) Family LONDON’S GLOBAL UNIVERSITY Analysis of the novel Lipid transfer protein Anchored at Membrane contact sites (LAM) family A thesis submitted to the University College London in the fulfilment of the requirements for the degree of Doctor of Philosophy Supervisor: Dr Tim Levine Department of Cell Biology, Faculty of Brain Science, Institute of Ophthalmology 2017 Declaration I, Louise Wong, confirm that the work presented in this thesis is my own. Where information has been derived from other sources, I confirm that this has been indicated. June 2017 1 2 Abstract Membrane contact sites are dynamic structures where two organelles come into close proximity to regulate and facilitate the flow of material and information between them. One type of inter-organelle communication is lipid exchange, which is essential for membrane maintenance and in response to environmental and cellular stimuli. We recently discovered a new family of Lipid transfer proteins Anchored at Membrane contact sites (LAMs) that is present in all eukaryotes. LAM proteins are integral Endoplasmic Reticulum (ER) proteins containing at least one domain that is structurally similar to the StARkin domain superfamily, a specialised fold that can bind amphipathic ligands such as lipids. The budding yeast, Saccharomyces cerevisiae, has six such proteins: Lam1p-6p. Lam1p-4p are located at contacts between the ER and the plasma membrane (PM), and Lam1p-3p are implicated in retrograde sterol traffic between the ER and PM. The PM contains a high concentration of sterol where it increases rigidity by altering the packing characteristics of the phospholipids in the bilayer. Sterol is also important in the ER, where its levels are low but it is both synthesised and sensed. However, the mechanism by which sterol traffics between the ER and the PM is unknown. This investigation characterises the phenotype of yeast delete LAM strains on Amphotericin B, a sterol sequestering antifungal agent and shows that the conserved StARkin domain of LAM proteins is responsible for resistance against Amphotericin B. Aspergillus fumigatus, a filamentous fungus, has two LAM proteins and the removal of AfLamA causes a severe growth phenotype. Also, in vitro studies indicate that LAM StARkin domains have a clear sterol transfer activity and a mutation that can diminish the function in vivo and in vitro has been identified. These findings present a new candidate protein family for intracellular sterol trafficking. 3 4 Dedication To my beloved brother, Howard Wong, 1985-2012 I dedicate this thesis to: my brother, you are dearly missed and always remembered and to my mother, for your encouragement and infinite support. 5 6 Acknowledgements I would like to thank my supervisor, Dr Tim Levine, for the guidance and inspiration through what has been an exciting and at times, a challenging project. Your expertise, creativity and passion have helped me become the scientist that I am and aspire to be for the rest of my career. I could not have learnt or achieved as much as I did without your supervision and influence and most significantly your tremendous enthusiasm for all things science! I am extremely thankful that you gave me the chance to work on such an original project and more importantly as part of your lab. The members of the Levine group have contributed immensely to my work and to my time in the lab: first and foremost, Alberto: thanks for being there from the start. It was my pleasure to have worked together on this project and also undertaking the PhD journey with you. I have no doubts that sharing lab and office space with me was not always easy with my ‘organised’ mess, so thanks for putting up with me! Sarah: your infectious energy inspired me when I needed motivation. Rachel: thanks for showing the way to becoming a fully-fledged PhD student. My time at the Institute of Ophthalmology has been made easier by many staff members who maintain equipment and support all of us here, thank you for your help. I am also deeply indebted for the expert discussions, advice and resources to all the brilliant scientists from all around the world whom I have met over the course of my PhD. I gratefully acknowledge the funding from the MRC that made my PhD possible. I also extend my gratitude to the Cockcroft lab for the lipid transfer expertise, Cordeiro lab (UCL) and Ben who has helped made many experiments possible and also the F2G Ltd lab (Manchester) who supported both my PhD funding and a significant part of this project especially Nicola. To all fellow lab/office members, past and present, I thank you for being there through the ups and downs of experiments and life, providing support and most importantly friendship (and also so much conversation that I thought I would never get any work done!). I am especially grateful to Ingrid and Tom for being late night office buddies – I’m sure I would have been more motivated to finish my work quicker if it were not for your excellent company. To all friends and family, I thank you for being part of my life. Now this thesis is done and I am no longer a student, I definitely do not have any excuse for not sharing the responsibility at the pub. To my family, a huge thanks for all their love and encouragement. To my parents, I am so grateful that you gave me the opportunities to study that you did not 7 have for yourselves and for your extraordinary support you have always offered. I hope that I have made you proud. To my brothers, you have been there throughout my life and, you influenced so much of who I am, thus I could not have started this journey without you. Finally, my dearest Thomas, you have been by my side throughout as my unwavering rock through the toughest and the best of times giving me unconditional support, care and friendship. The completion of my PhD has been a long journey and life did not stand still during this time. It is only due to your extraordinary patience, kindness and love that I am able to be here. You encouraged me from afar when I first started this PhD and then you gave me strength when my brother passed away. We have begun our life together in London, and I can never be happier than coming home to you (and our cats!). When life was the difficult during my diagnosis, surgery and recovery and also the long time before that when I was not quite right, you never left me alone. You supported me mentally, emotionally and physically and we prevailed because of you. Writing is not my forte, but you have tolerated my complaints and whining during the completion of this thesis. There are no words that can express my gratitude and appreciation for all that you have done and been for me. This achievement is as much yours as mine. I thank you with all my heart and soul for being you. 8 Contents Declaration ..................................................................................................................... 1 Abstract .......................................................................................................................... 3 Dedication ...................................................................................................................... 5 Acknowledgements ........................................................................................................ 7 List of Figures .............................................................................................................. 17 List of Tables ............................................................................................................... 21 List of Abbreviations .................................................................................................. 23 CHAPTER 1 Introduction ......................................................................................... 29 1.1 General Introduction .................................................................................... 29 1.2 Lipids and cellular membranes ................................................................... 29 1.2.1 Lipid bilayers ................................................................................................ 29 1.2.2 Function of sterol in a membrane ................................................................. 31 1.2.3 Regulation of cholesterol synthesis .............................................................. 34 1.2.4 Intracellular distribution of sterol ................................................................. 36 1.2.5 Sterol affinity in membranes ........................................................................ 37 1.2.6 Sterol pools ................................................................................................... 38 1.3 Intracellular lipid transfer ........................................................................... 38 1.3.1 Three classes of intracellular lipid movement .............................................. 38 1.3.2 Transport of cellular sterol between membranes .......................................... 39 1.3.3 Complications of unravelling intracellular sterol transport .......................... 40 1.3.4 Vesicular pathway is not responsible for bulk sterol transport .................... 41 1.3.5 Spontaneous diffusion is not sufficient ........................................................ 42 1.3.6 Lipid transfer by proteins at contact sites ..................................................... 43 1.4 Membrane Contact Sites .............................................................................. 44 1.4.1 ER membrane contact sites..........................................................................
Load more

Recommended publications
  • Predicting Protein-Membrane Interfaces of Peripheral Membrane
    bioRxiv preprint doi: https://doi.org/10.1101/2021.06.28.450157; this version posted June 29, 2021. The copyright holder for this preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under aCC-BY-NC-ND 4.0 International license. Predicting protein-membrane interfaces of pe- ripheral membrane proteins using ensemble machine learning Alexios Chatzigoulas1,2,* and Zoe Cournia1,* 1Biomedical Research Foundation, Academy of Athens, 4 Soranou Ephessiou, 11527 Athens, Greece, 2Depart- ment of Informatics and Telecommunications, National and Kapodistrian University of Athens, 15784 Athens, Greece *To whom correspondence should be addressed. Abstract Motivation: Abnormal protein-membrane attachment is involved in deregulated cellular pathways and in disease. Therefore, the possibility to modulate protein-membrane interactions represents a new promising therapeutic strategy for peripheral membrane proteins that have been considered so far undruggable. A major obstacle in this drug design strategy is that the membrane binding domains of peripheral membrane proteins are usually not known. The development of fast and efficient algorithms predicting the protein-membrane interface would shed light into the accessibility of membrane-protein interfaces by drug-like molecules. Results: Herein, we describe an ensemble machine learning methodology and algorithm for predicting membrane-penetrating residues. We utilize available experimental data in the literature for training 21 machine learning classifiers and a voting classifier. Evaluation of the ensemble classifier accuracy pro- duced a macro-averaged F1 score = 0.92 and an MCC = 0.84 for predicting correctly membrane-pen- etrating residues on unknown proteins of an independent test set.
    [Show full text]
  • An Overview of Lipid Membrane Models for Biophysical Studies
    biomimetics Review Mimicking the Mammalian Plasma Membrane: An Overview of Lipid Membrane Models for Biophysical Studies Alessandra Luchini 1 and Giuseppe Vitiello 2,3,* 1 Niels Bohr Institute, University of Copenhagen, Universitetsparken 5, 2100 Copenhagen, Denmark; [email protected] 2 Department of Chemical, Materials and Production Engineering, University of Naples Federico II, Piazzale Tecchio 80, 80125 Naples, Italy 3 CSGI-Center for Colloid and Surface Science, via della Lastruccia 3, 50019 Sesto Fiorentino (Florence), Italy * Correspondence: [email protected] Abstract: Cell membranes are very complex biological systems including a large variety of lipids and proteins. Therefore, they are difficult to extract and directly investigate with biophysical methods. For many decades, the characterization of simpler biomimetic lipid membranes, which contain only a few lipid species, provided important physico-chemical information on the most abundant lipid species in cell membranes. These studies described physical and chemical properties that are most likely similar to those of real cell membranes. Indeed, biomimetic lipid membranes can be easily prepared in the lab and are compatible with multiple biophysical techniques. Lipid phase transitions, the bilayer structure, the impact of cholesterol on the structure and dynamics of lipid bilayers, and the selective recognition of target lipids by proteins, peptides, and drugs are all examples of the detailed information about cell membranes obtained by the investigation of biomimetic lipid membranes. This review focuses specifically on the advances that were achieved during the last decade in the field of biomimetic lipid membranes mimicking the mammalian plasma membrane. In particular, we provide a description of the most common types of lipid membrane models used for biophysical characterization, i.e., lipid membranes in solution and on surfaces, as well as recent examples of their Citation: Luchini, A.; Vitiello, G.
    [Show full text]
  • Membrane Proteins Are Associated with the Membrane of a Cell Or Particular Organelle and Are Generally More Problematic to Purify Than Water-Soluble Proteins
    Strategies for the Purification of Membrane Proteins Sinéad Marian Smith Department of Clinical Medicine, School of Medicine, Trinity College Dublin, Ireland. Email: [email protected] Abstract Although membrane proteins account for approximately 30 % of the coding regions of all sequenced genomes and play crucial roles in many fundamental cell processes, there are relatively few membranes with known 3D structure. This is likely due to technical challenges associated with membrane protein extraction, solubilization and purification. Membrane proteins are classified based on the level of interaction with membrane lipid bilayers, with peripheral membrane proteins associating non- covalently with the membrane, and integral membrane proteins associating more strongly by means of hydrophobic interactions. Generally speaking, peripheral membrane proteins can be purified by milder techniques than integral membrane proteins, whose extraction require phospholipid bilayer disruption by detergents. Here, important criteria for strategies of membrane protein purification are addressed, with a focus on the initial stages of membrane protein solublilization, where problems are most frequently are encountered. Protocols are outlined for the successful extraction of peripheral membrane proteins, solubilization of integral membrane proteins, and detergent removal which is important not only for retaining native protein stability and biological functions, but also for the efficiency of downstream purification techniques. Key Words: peripheral membrane protein, integral membrane protein, detergent, protein purification, protein solubilization. 1. Introduction Membrane proteins are associated with the membrane of a cell or particular organelle and are generally more problematic to purify than water-soluble proteins. Membrane proteins represent approximately 30 % of the open-reading frames of an organism’s genome (1-4), and play crucial roles in basic cell functions including signal transduction, energy production, nutrient uptake and cell-cell communication.
    [Show full text]
  • The Structure of Biological Membranes
    The Structure of Biological Membranes Func7ons of Cellular Membranes 1. Plasma membrane acts as a selecvely permeable barrier to the environment • Uptake of nutrients • Waste disposal • Maintains intracellular ionic milieau 2. Plasma membrane facilitates communicaon • With the environment • With other cells • Protein secreon 3. Intracellular membranes allow compartmentalizaon and separaon of different chemical reac7on pathways • Increased efficiency through proximity • Prevent fu8le cycling through separaon Composi7on of Animal Cell Membranes • Hydrated, proteinaceous lipid bilayers • By weight: 20% water, 80% solids • Solids: Lipid Protein ~90% Carbohydrate (~10%) • Phospholipids responsible for basic membrane bilayer structure and physical proper8es • Membranes are 2-dimensional fluids into which proteins are dissolved or embedded The Most Common Class of PhospholipiD is FormeD from a Gycerol-3-P Backbone SaturateD FaJy AciD •Palmitate and stearate most common •14-26 carbons •Even # of carbons UnsaturateD FaJy AciD Structure of PhosphoglyceriDes All Membrane LipiDs are Amphipathic Figure 10-2 Molecular Biology of the Cell (© Garland Science 2008) PhosphoglyceriDes are Classified by their Head Groups Phosphadylethanolamine Phosphadylcholine Phosphadylserine Phosphadylinositol Ether Bond at C1 PS and PI bear a net negave charge at neutral pH Sphingolipids are the SeconD Major Class of PhospholipiD in Animal Cells Sphingosine Ceramides contain sugar moies in ether linkage to sphingosine GlycolipiDs are AbunDant in Brain Cells Figure 10-18 Molecular
    [Show full text]
  • Biosynthesis of New Alpha-Bisabolol Derivatives Through a Synthetic Biology Approach Arthur Sarrade-Loucheur
    Biosynthesis of new alpha-bisabolol derivatives through a synthetic biology approach Arthur Sarrade-Loucheur To cite this version: Arthur Sarrade-Loucheur. Biosynthesis of new alpha-bisabolol derivatives through a synthetic biology approach. Biochemistry, Molecular Biology. INSA de Toulouse, 2020. English. NNT : 2020ISAT0003. tel-02976811 HAL Id: tel-02976811 https://tel.archives-ouvertes.fr/tel-02976811 Submitted on 23 Oct 2020 HAL is a multi-disciplinary open access L’archive ouverte pluridisciplinaire HAL, est archive for the deposit and dissemination of sci- destinée au dépôt et à la diffusion de documents entific research documents, whether they are pub- scientifiques de niveau recherche, publiés ou non, lished or not. The documents may come from émanant des établissements d’enseignement et de teaching and research institutions in France or recherche français ou étrangers, des laboratoires abroad, or from public or private research centers. publics ou privés. THÈSE En vue de l’obtention du DOCTORAT DE L’UNIVERSITÉ DE TOULOUSE Délivré par l'Institut National des Sciences Appliquées de Toulouse Présentée et soutenue par Arthur SARRADE-LOUCHEUR Le 30 juin 2020 Biosynthèse de nouveaux dérivés de l'α-bisabolol par une approche de biologie synthèse Ecole doctorale : SEVAB - Sciences Ecologiques, Vétérinaires, Agronomiques et Bioingenieries Spécialité : Ingénieries microbienne et enzymatique Unité de recherche : TBI - Toulouse Biotechnology Institute, Bio & Chemical Engineering Thèse dirigée par Gilles TRUAN et Magali REMAUD-SIMEON Jury
    [Show full text]
  • The Membrane Interface of Chloroplast Signal Recognition Particle-Dependent Protein Targeting
    University of Arkansas, Fayetteville ScholarWorks@UARK Theses and Dissertations 5-2009 The eM mbrane Interface of Chloroplast Signal Recognition Particle-Dependent Protein Targeting Naomi Jane Marty University of Arkansas, Fayetteville Follow this and additional works at: http://scholarworks.uark.edu/etd Part of the Cell Biology Commons, and the Molecular Biology Commons Recommended Citation Marty, Naomi Jane, "The eM mbrane Interface of Chloroplast Signal Recognition Particle-Dependent Protein Targeting" (2009). Theses and Dissertations. 6. http://scholarworks.uark.edu/etd/6 This Dissertation is brought to you for free and open access by ScholarWorks@UARK. It has been accepted for inclusion in Theses and Dissertations by an authorized administrator of ScholarWorks@UARK. For more information, please contact [email protected], [email protected]. THE MEMBRANE INTERFACE OF CHLOROPLAST SIGNAL RECOGNITION PARTICLE-DEPENDENT PROTEIN TARGETING THE MEMBRANE INTERFACE OF CHLOROPLAST SIGNAL RECOGNITION PARTICLE-DEPENDENT PROTEIN TARGETING A dissertation submitted in partial fulfillment of the requirements for the degree of Doctor of Philosophy in Cell and Molecular Biology By Naomi J. Marty Northeastern State University Bachelor of Science in Biology, 2003 May 2009 University of Arkansas ABSTRACT A novel signal recognition particle (SRP) found in the chloroplast (cpSRP) works in combination with the cpSRP receptor, cpFtsY, to facilitate the post-translational targeting of a family of nuclear-encoded thylakoid proteins to the Alb3 translocase in thylakoid membranes. Work here focused on understanding events at the membrane that take place to ensure targeting of the cpSRP-dependent substrate to Alb3. Specifically, we sought to understand the structural and functional role of membrane binding by cpFtsY, a protein that exhibits the ability to partition between the membrane (thylakoid) and soluble (stroma) phase during protein targeting.
    [Show full text]
  • The Lipid Bilayer: Composition and Structural Organization
    THE LIPID BILAYER: COMPOSITION AND STRUCTURAL ORGANIZATION • MR. SOURAV BARAI • ASSISTANT PROFESSOR • DEPARTMENT OF ZOOLOGY • JHARGRAM RAJ COLLEGE THELIPID BILAYER: COMPOSITION AND STRUCTURAL ORGANIZATION The Fluid Mosaic Model of Biomembrane Plasma membrane • 1. Affect shape and function • 2. Anchor protein to the membrane • 3. Modify membrane protein activities • 4. Transducing signals to the cytoplasm “A living cell is a self-reproducing system of molecules held inside a container - the plasma membrane” Membrane comprised of lipid sheet (5 nm thick) • Primary purpose - barrier to prevent cell contents spilling out BUT, must be selective barrier Lipid Composition and struCturaL organization • Phospholipids of the composition present in cells spontaneously form sheet like phospholipid bilayers, which are two molecules thick. • The hydrocarbon chains of the phospholipids in each layer, or leaflet, form a hydrophobic core that is 3–4 nm thick in most biomembranes. • Approx 10^6 lipid molecule in 1µm×1µm area of lipid bilayer. • Electron microscopy of thin membrane sections stained with osmium tetroxide, which binds strongly to the polar head groups of phospholipids, reveals the bilayer structure. • A cross section of all single membranes stained with osmium tetroxide looks like a railroad track: two thin dark lines (the stain–head group complexes) with a uniform light space of about 2nm (the hydrophobic tails) between them. PROPERTIES • PERMIABILITY: The hydrophobic core is an impermeable barrier that prevents the diffusion of water-soluble (hydrophilic) solutes across the membrane. • STABILITY: The bilayer structure is maintained by hydrophobic and van der Waals interactions between the lipid chains. Even though the exterior aqueous environment can vary widely in ionic strength and pH, the bilayer has the strength to retain its characteristic architecture.
    [Show full text]
  • ERG11) Gene of Moniliophthora Perniciosa
    Genetics and Molecular Biology, 37, 4, 683-693 (2014) Copyright © 2014, Sociedade Brasileira de Genética. Printed in Brazil www.sbg.org.br Research Article Analysis of the ergosterol biosynthesis pathway cloning, molecular characterization and phylogeny of lanosterol 14 a-demethylase (ERG11) gene of Moniliophthora perniciosa Geruza de Oliveira Ceita1,4, Laurival Antônio Vilas-Boas2, Marcelo Santos Castilho3, Marcelo Falsarella Carazzolle5, Carlos Priminho Pirovani6, Alessandra Selbach-Schnadelbach4, Karina Peres Gramacho7, Pablo Ivan Pereira Ramos4, Luciana Veiga Barbosa4, Gonçalo Amarante Guimarães Pereira5 and Aristóteles Góes-Neto1 1Laboratório de Pesquisa em Microbiologia, Departamento de Ciências Biológicas, Universidade Estadual de Feira de Santana, Feira de Santana, BA, Brazil. 2Centro de Ciências Biológicas, Departamento de Biologia Geral, Universidade Estadual de Londrina, Londrina, PR, Brazil. 3Laboratório de Bioinformática e Modelagem Molecular, Departamento do Medicamento, Faculdade de Farmácia, Universidade Federal da Bahia, Salvador, BA, Brazil. 4Laboratório de Biologia Molecular, Instituto de Biologia, Departamento de Biologia Geral, Universidade Federal da Bahia, Salvador, BA, Brazil. 5Laboratório de Genômica e Proteômica, Departamento de Genética e Evolução, Universidade Estadual de Campinas, Campinas, SP, Brazil. 6Centro de Biotecnologia e Genética, Departamento de Ciências Biológicas, Universidade Estadual de Santa Cruz, Ilhéus, BA, Brazil. 7Laboratório de Fitopatologia Molecular, Centro de Pesquisas do Cacau, Ilhéus, BA, Brazil. Abstract The phytopathogenic fungus Moniliophthora perniciosa (Stahel) Aime & Philips-Mora, causal agent of witches’ broom disease of cocoa, causes countless damage to cocoa production in Brazil. Molecular studies have attempted to identify genes that play important roles in fungal survival and virulence. In this study, sequences deposited in the M. perniciosa Genome Sequencing Project database were analyzed to identify potential biological targets.
    [Show full text]
  • 85224601.Pdf
    View metadata, citation and similar papers at core.ac.uk brought to you by CORE provided by RERO DOC Digital Library RESEARCH LETTER Construction and characterization of Enterococcus faecalis CG110/gfp/pRE25Ã, a tool for monitoring horizontal gene transfer in complex microbial ecosystems Martina C. Haug, Sabine A. Tanner, Christophe Lacroix, Leo Meile & Marc J.A. Stevens Laboratory of Food Biotechnology, Institute of Food, Nutrition and Health, ETH Zurich, Zurich, Switzerland Correspondence: Leo Meile, Laboratory of Abstract Food Biotechnology, Institute of Food, Nutrition and Health, ETH Zurich, Enterococci are among the most notorious bacteria involved in the spread of Schmelzbergstrasse 7, CH-8092 Zurich, antibiotic resistance (ABR) determinants via horizontal gene transfer, a process that Switzerland. Tel.: 141 44 632 33 62; fax: leads to increased prevalence of antibiotic-resistant bacteria. In complex microbial 141 44 632 14 03; e-mail: communities with a high background of ABR genes, detection of gene transfer is [email protected] possible only when the ABR determinant is marked. Therefore, the conjugative multiresistance plasmid pRE25, originating from a sausage-associated Enterococcus Received 6 August 2010; revised 8 September faecalis, was tagged with a 34-bp random sequence marker spliced by tet(M). The 2010; accepted 29 September 2010. plasmid constructed, designated pRE25Ã, was introduced into E. faecalis CG110/ Final version published online 28 October 2010. gfp, a strain containing a gfp gene as chromosomal marker. The plasmid pRE25Ã is DOI:10.1111/j.1574-6968.2010.02131.x fully functional compared with its parental pRE25, occurs at one to two copies per chromosome, and can be transferred to Listeria monocytogenes and Listeria innocua À6 À8 Editor: Andre´ Klier at frequencies of 6 Â 10 to 8 Â 10 transconjugants per donor.
    [Show full text]
  • Klinicky Významné Rezistentní Plazmidy – Studium Biologické Zátěže a Stability Úspěšných Mobilních Genetických Elementů
    MASARYKOVA UNIVERZITA PŘÍRODOVĚDECKÁ FAKULTA ÚSTAV EXPERIMENTÁLNÍ BIOLOGIE Klinicky významné rezistentní plazmidy – studium biologické zátěže a stability úspěšných mobilních genetických elementů Diplomová práce Tomáš Nohejl Vedoucí práce: doc. RNDr. Monika Dolejská, Ph.D. Brno 2018 Bibliografický záznam Autor: Bc. Tomáš Nohejl Přírodovědecká fakulta, Masarykova univerzita Ústav experimentální biologie Název práce: Klinicky významné rezistentní plazmidy – studium biologické zátěže a stability úspěšných mobilních genetických elementů Studijní program: Experimentální biologie Studijní obor: Molekulární biologie a genetika Vedoucí práce: doc. RNDr. Monika Dolejská, Ph.D. Akademický rok: 2017/2018 Počet stran 78 Klíčová slova: Antibiotická rezistence; plazmid; horizontální přenos genetické informace; HGT; konjugace; PMQR; chinolony; fitness; perzistence Bibliographic entry Author: Bc. Tomáš Nohejl Faculty of Science, Masaryk University Department of Experimental Biology Title of Thesis: Clinically important resistance plasmids - the study of biological burden and stability of successful mobile genetic elements Degree programme: Experimental biology Field of Study: Molecular Biology and Genetics Supervisor: doc. RNDr. Monika Dolejská, Ph.D. Academic Year: 2017/2018 Number of Pages: 78 Keywords: Antibiotic resistance; plasmid; horizontal gene transfer; HGT; conjugation; PMQR; quinolones; fitness; persistence Abstrakt Antibiotická rezistence představuje jeden z nejzávažnějších globálních problémů současnosti. Šíření genů kódující antibiotickou
    [Show full text]
  • A Combined Growth Factor-Deleted and Thymidine Kinase-Deleted Vaccinia Virus Vector
    (19) & (11) EP 2 325 321 A1 (12) EUROPEAN PATENT APPLICATION (43) Date of publication: (51) Int Cl.: 25.05.2011 Bulletin 2011/21 C12N 15/863 (2006.01) A61K 48/00 (2006.01) (21) Application number: 10179286.9 (22) Date of filing: 26.05.2000 (84) Designated Contracting States: • Bartlett, David L. AT BE CH CY DE DK ES FI FR GB GR IE IT LI LU Darnestown, MD 20878 (US) MC NL PT SE • Moss, Bernard Bethesda, MD 20814 (US) (30) Priority: 28.05.1999 US 137126 P (74) Representative: Donald, Jenny Susan (62) Document number(s) of the earlier application(s) in Forrester & Boehmert accordance with Art. 76 EPC: Pettenkoferstrasse 20-22 00939374.5 / 1 180 157 80336 München (DE) (71) Applicant: THE GOVERNMENT OF THE UNITED Remarks: STATES OF AMERICA as •This application was filed on 24-09-2010 as a represented by the SECRETARY OF THE divisional application to the application mentioned DEPARTMENT OF under INID code 62. HEALTH AND HUMAN SERVICES •Claims filed after the date of filing of the application Rockville, MD 20852 (US) / after the date of receipt of the divisional appliaction (Rule 68(4) EPC). (72) Inventors: • McCart, Andrea J. Silver Springs, MD 20910 (US) (54) A combined growth factor-deleted and thymidine kinase-deleted vaccinia virus vector (57) A composition of matter comprising a vaccinia virus expression vector with a negative thymidine kinase phe- notype and a negative vaccinia virus growth factor phenotype. EP 2 325 321 A1 Printed by Jouve, 75001 PARIS (FR) EP 2 325 321 A1 Description Background of the Invention 5 Field of the Invention [0001] The present invention relates to mutant vaccinia virus expression vectors The mutant expression vectors of the present invention show substantially no virus replication in non dividing cells and as such are superior to previous vaccinia virus expression vectors.
    [Show full text]
  • A Phosphorylated Transcription Factor Regulates Sterol Biosynthesis in Fusarium Graminearum
    ARTICLE https://doi.org/10.1038/s41467-019-09145-6 OPEN A phosphorylated transcription factor regulates sterol biosynthesis in Fusarium graminearum Zunyong Liu1,2, Yunqing Jian1,2, Yun Chen 1,2, H. Corby Kistler 3, Ping He4, Zhonghua Ma 1,2 & Yanni Yin1,2 Sterol biosynthesis is controlled by transcription factor SREBP in many eukaryotes. Here, we show that SREBP orthologs are not involved in the regulation of sterol biosynthesis in Fusarium graminearum, a fungal pathogen of cereal crops worldwide. Instead, sterol produc- 1234567890():,; tion is controlled in this organism by a different transcription factor, FgSR, that forms a homodimer and binds to a 16-bp cis-element of its target gene promoters containing two conserved CGAA repeat sequences. FgSR is phosphorylated by the MAP kinase FgHog1, and the phosphorylated FgSR interacts with the chromatin remodeling complex SWI/SNF at the target genes, leading to enhanced transcription. Interestingly, FgSR orthologs exist only in Sordariomycetes and Leotiomycetes fungi. Additionally, FgSR controls virulence mainly via modulating deoxynivalenol biosynthesis and responses to phytoalexin. 1 State Key Laboratory of Rice Biology, Zhejiang University, 866 Yuhangtang Road, Hangzhou 310058, China. 2 Institute of Biotechnology, Key Laboratory of Molecular Biology of Crop Pathogens and Insects, Zhejiang University, 866 Yuhangtang Road, Hangzhou 310058, China. 3 United States Department of Agriculture, Agricultural Research Service, 1551 Lindig Street, St. Paul, MN 55108, USA. 4 Department of Biochemistry
    [Show full text]