DOCSLIB.ORG

Use of Stomatin (Stm1) Polynucleotides For

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Use of Stomatin (Stm1) Polynucleotides For (19) TZZ__T (11) EP 1 979 484 B1 (12) EUROPEAN PATENT SPECIFICATION (45) Date of publication and mention (51) Int Cl.: of the grant of the patent: C12N 15/85 (2006.01) C07K 14/425 (2006.01) 19.03.2014 Bulletin 2014/12 A01H 5/00 (2006.01) (21) Application number: 07703625.9 (86) International application number: PCT/EP2007/050062 (22) Date of filing: 04.01.2007 (87) International publication number: WO 2007/080143 (19.07.2007 Gazette 2007/29) (54) USE OF STOMATIN (STM1) POLYNUCLEOTIDES FOR ACHIEVING A PATHOGEN RESISTANCE IN PLANTS VERWENDUNG VON STOMATIN (STM1)-POLYNUKLEOTIDEN ZUR ERREICHUNG EINER RESISTENZ GEGEN KRANKHEITSERREGER VON PFLANZEN UTILISATION DE POLYNUCLÉOTIDES DE LA STOMATINE (STM1) POUR OBTENIR UNE RÉSISTANCE À UN PATHOGÈNE DANS DES VÉGÉTAUX (84) Designated Contracting States: • NADIMPALLI RAMGOPAL ET AL: "Prohibitins, AT BE BG CH CY CZ DE DK EE ES FI FR GB GR stomatins, and plant disease response genes HU IE IS IT LI LT LU LV MC NL PL PT RO SE SI compose a protein superfamily that controls cell SK TR proliferation, ion channel regulation, and death" JOURNAL OF BIOLOGICAL CHEMISTRY, vol. (30) Priority: 12.01.2006 EP 06100304 275, no. 38, 22 September 2000 (2000-09-22), pages 29579-29586, XP002431766 ISSN: (43) Date of publication of application: 0021-9258 15.10.2008 Bulletin 2008/42 • OPALSKI KRYSTINA S ET AL: "The receptor- like MLO protein and the RAC/ROP family G-protein (60) Divisional application: RACB modulate actin reorganization in barley 11163021.6 / 2 380 986 attacked by the biotrophic powdery mildew fungus Blumeria graminis f.sp hordei" PLANT (73) Proprietor: BASF Plant Science GmbH JOURNAL, vol. 41, no. 2, January 2005 (2005-01), 67056 Ludwigshafen (DE) pages 291-303, XP002431767 ISSN: 0960-7412 • BHAT RIYAZ A ET AL: "Recruitment and (72) Inventors: interaction dynamics of plant penetration • FRANK, Markus resistance components in a plasma membrane 67434 Neustadt (DE) microdomain" PROCEEDINGS OF THE • SCHWEIZER, Patrick NATIONAL ACADEMY OF SCIENCES OF THE 06466 Gatersleben (DE) UNITED STATES OF AMERICA, vol. 102, no. 8, 22 • DOUCHKOV, Dimitar February 2005 (2005-02-22), pages 3135-3140, 06466 Gatersleben (DE) XP002431768 ISSN: 0027-8424 • HUECKELHOVEN R ET AL: "Overexpression of (74) Representative: Popp, Andreas barley BAX inhibitor 1 induces breakdown of mlo- BASF SE mediated penetration resistance to Blumeria Global Intellectual Property graminis" PROCEEDINGS OF THE NATIONAL GVX - C6 ACADEMY OF SCIENCES OF USA, NATIONAL 67056 Ludwigshafen (DE) ACADEMY OF SCIENCE, WASHINGTON, DC, US, vol. 100, no. 9, 29 April 2003 (2003-04-29), pages (56) References cited: 5555-5560, XP002286823 ISSN: 0027-8424 WO-A-00/15817 WO-A-03/020939 Note: Within nine months of the publication of the mention of the grant of the European patent in the European Patent Bulletin, any person may give notice to the European Patent Office of opposition to that patent, in accordance with the Implementing Regulations. Notice of opposition shall not be deemed to have been filed until the opposition fee has been paid. (Art. 99(1) European Patent Convention). EP 1 979 484 B1 Printed by Jouve, 75001 PARIS (FR) (Cont. next page) EP 1 979 484 B1 • HAMMOND-KOSACK K E ET AL: "RESISTANCE GENE-DEPENDENT PLANT DEFENSE RESPONSES" PLANT CELL, AMERICAN SOCIETY OF PLANT PHYSIOLOGISTS, ROCKVILLE, MD, US, vol. 8, no. 10, October 1996 (1996-10), pages 1773-1791, XP002049471 ISSN: 1040-4651 2 EP 1 979 484 B1 Description [0001] The invention relates to a method of generating or increasing a pathogen resistance in plants by reducing the expression of at least one stomatin polypeptide or a functional equivalent thereof. The invention relates to novel nucleic 5 acid sequences coding for a Hordeum vulgare stomatin (HvSTM1) polynucleotide and describes homologous sequences (STM1) thereof, and to their use in methods for obtaining a pathogen resistance in plants, and to nucleic acid constructs, expression cassettes and vectors which comprise these sequences and which are suitable for mediating a fungal re- sistance in plants. The invention furthermore relates to transgenic organisms, in particular plants, which are transformed with these expression cassettes or vectors, and to cultures, parts or transgenic propagation material derived therefrom. 10 [0002] There are only few approaches which confer a resistance to pathogens, mainly fungal pathogens, to plants. This shortcoming can partly be attributed to the complexity of the biological systems in question. Another fact which stands in the way of obtaining resistances to pathogens is that little is known about the interactions between pathogen and plant. The large number of different pathogens, the infection mechanisms developed by these organisms and the defence mechanisms developed by the plant phyla, families and species interact with one another in many different ways. 15 [0003] Fungal pathogens have developed essentially two infection strategies. Some fungi enter into the host tissue via the stomata (for example rusts, Septoria species, Fusarium species) and penetrate the mesophyll tissue, while others penetrate via the cuticles into the epidermal cells underneath (for example Blumeria species). [0004] The infections caused by the fungal pathogens lead to the activation of the plant’s defence mechanisms in the infected plants. Thus, it has been possible to demonstrate that defence reactions against epidermis-penetrating fungi 20 frequently start with the formation of a penetration resistance (formation of papillae, strengthening of the cell wall with callose as the main constituent) underneath the fungal penetration hypha (Elliott et al. Mol Plant Microbe Interact. 15: 1069-77; 2002). [0005] In some cases, however, the plant’s defence mechanisms only confer an insufficient protection mechanism against the attack by pathogens. 25 [0006] The formation of a penetration resistance to pathogens whose infection mechanism comprises a penetration of the epidermal cells or of the mesophyll cells is of great importance both for monocotyledonous and for dicotyledonous plants. In contrast to described mlo-mediated resistance, it can probably make possible the development of a broad- spectrum resistance against obligatory biotrophic, hemibiotrophic and necrotrophic fungi. [0007] The present invention was therefore based on the object of providing a method for generating a resistance of 30 plants to penetrating pathogens. [0008] The object is achieved by the embodiments characterized in the claims. [0009] The invention therefore relates to a method of increasing the resistance to penetrating pathogens in a mono- cotyledonous or dicotyledonous plant, or a part of a plant, for example in an organ, tissue, a cell or a part of a plant cell, for example in an organelle, which comprises lessening or reducing the activity of a stomatin protein according to SEQ 35 ID No 2 (STM1) in the plant, or a part of the plant, for example in an organ, tissue, a cell or a part of a cell, for example in a cell compartment, for example in an organelle, in comparison with a control plant or a part of a control plant, for example its organ, tissue, cell or part of a cell, for example in a cell compartment, for example in an organelle. [0010] Preferably, a race- unspecific resistance is obtained in the method according to the invention. Thus, for example, a broad-spectrum resistance against obligatorily biotrophic and/or hembiotrophic and/or necrotrophic fungi of plants, in 40 particular against mesophyll-penetrating pathogens, can be obtained by the method according to the invention. [0011] Surprisingly, it has been observed that the gene silencing via dsRNAi of a gene which codes for the stomatin protein HvSTM1 results in an increase in the resistance of monocotyledonous and dicotyledonous plants to fungal pathogens. Thus, this negative control function in the event of attack by fungal pathogens has been demonstrated for the stomatin protein HvSTM1 from barley (Hordeum vulgare) (HvSTM1), wheat (Triticum aestivum) and thale cress 45 (Arabidopsis thaliana). [0012] It has been found within the scope of a TIGS (=Transient Induced Gene Silencing) analysis in barley by the method of Schweizer et al. (2001) that a dsRNAi- mediated silencing of the gene HvSTM greatly increases the resistance to Blumeria graminis f. sp. hordei (synonym: Erysiphe graminis DC. f. sp. hordei). This effect has also been obtained in dicotyledonous species such as, for example, Arabidopsis thaliana by inducing the post-transcriptional gene silencing 50 (PTGS). This emphasizes the universal importance of the loss- of-function of HvSTM1-homologous genes for the devel- opment of a broad-spectrum pathogen resistance of the plant. [0013] Stomatin is an integral membrane protein which has first been identified in blood cells. In certain hereditary diseases, in which this protein is absent, a hemolytic anemia results. In this stomatocytosis, the blood cells suffer a pronounced passive diffusion of singly charged cations, which results in hyperhydration as the result of a high sodium 55 concentration and a low potassium concentration. The name stomatin is derived from the mouth- like structure of these erythrocytes (Greek Stoma = mouth). Stomatin acts as a negative membrane permeability regulator for singly charged cations. Stomatin has a single membrane domain, while the remainder of the protein protrudes into the cytoplasm. The molecular mechanisms via which stomatin exerts its function is unclear. It is assumed that the cytoplasmic domain of 3 EP 1 979 484 B1 the protein acts sterically as a sort of plug which closes ion channels and perhaps interacts with the cytoskeleton (Stewart GW, Argent AC, Dash BCJ (1993) Biochim. Biophys. Acta 1225, 15-25; Stewart GW et al. (1992) Blood 79, 1593-1601). [0014] A first potential plant stomatin was cloned in maize (Nadimpalli R et al. (2000) J. Biol. Chem. 275, 29579-29586). However, no functional data for this gene, Zm-stm1, were published. A potential role in pathogen defence has been 5 assumed for other genes which were cloned in this work since transcripts of these genes were upregulated in the lesion Mimic-Mutante Les9.
Load more

Recommended publications
  • Mechanistic Insights Into GLUT1 Activation and Clustering Revealed by Super-Resolution Imaging
    Mechanistic insights into GLUT1 activation and clustering revealed by super-resolution imaging Qiuyan Yana,b, Yanting Lub,c, Lulu Zhoua,b, Junling Chena, Haijiao Xua, Mingjun Caia, Yan Shia, Junguang Jianga, Wenyong Xiongc,1, Jing Gaoa,1, and Hongda Wanga,d,1 aState Key Laboratory of Electroanalytical Chemistry, Research Center of Biomembranomics, Changchun Institute of Applied Chemistry, Chinese Academy of Sciences, Changchun, 130022 Jilin, P. R. China; bSchool of Chemistry and Chemical Engineering, University of Chinese Academy of Sciences, 100049 Beijing, P. R. China; cState Key Laboratory of Phytochemistry and Plant Resources in West China, Kunming Institute of Botany, Chinese Academy of Sciences, Kunming, 650201 Yunnan, P. R. China; and dLaboratory for Marine Biology and Biotechnology, Qingdao National Laboratory for Marine Science and Technology, Aoshanwei, Jimo, Qingdao, 266237 Shandong, P. R. China Edited by Nieng Yan, Princeton University, Princeton, NJ, and accepted by Editorial Board Member Alan R. Fersht May 24, 2018 (received for review March 9, 2018) The glucose transporter GLUT1, a plasma membrane protein that super-resolution fluorescence microscopy, which breaks the dif- mediates glucose homeostasis in mammalian cells, is responsible fraction barrier and achieves a lateral resolution in the tens of for constitutive uptake of glucose into many tissues and organs. nanometers (17), has provided a particularly suitable tool to solve Many studies have focused on its vital physiological functions and these problems. Meanwhile, it has been proven that multiprotein close relationship with diseases. However, the molecular mecha- assemblies are dependent on cholesterol environment, and their nisms of its activation and transport are not clear, and its detailed separation and anchoring are related to the actin cytoskeleton (18, distribution pattern on cell membranes also remains unknown.
    [Show full text]
  • NDUFAB1 Protects Heart by Coordinating Mitochondrial Respiratory Complex
    bioRxiv preprint doi: https://doi.org/10.1101/302281; this version posted April 16, 2018. The copyright holder for this preprint (which was not certified by peer review) is the author/funder. All rights reserved. No reuse allowed without permission. NDUFAB1 Protects Heart by Coordinating Mitochondrial Respiratory Complex and Supercomplex Assembly Running title: Hou et al. Cardiac Protection by NDUFAB1 Tingting Hou 1; Rufeng Zhang 1; Chongshu Jian 1; Wanqiu Ding 1; Yanru Wang 1; Qi Ma 1; Xinli Hu 1; Heping Cheng 1,†; Xianhua Wang 1,† 1State Key Laboratory of Membrane Biology, Beijing Key Laboratory of Cardiometabolic Molecular Medicine, Peking-Tsinghua Center for Life Sciences, Institute of Molecular Medicine, Peking University, China. † Corresponding author. Xianhua Wang Tel: 86-10-62754605 Email: [email protected] Heping Cheng Tel: 86-10-62765957 Email: [email protected] bioRxiv preprint doi: https://doi.org/10.1101/302281; this version posted April 16, 2018. The copyright holder for this preprint (which was not certified by peer review) is the author/funder. All rights reserved. No reuse allowed without permission. Abstract The impairment of mitochondrial bioenergetics, often coupled with exaggerated reactive oxygen species (ROS) production, is emerging as a common mechanism in diseases of organs with a high demand for energy, such as the heart. Building a more robust cellular powerhouse holds promise for protecting these organs in stressful conditions. Here, we demonstrate that NDUFAB1 (NADH:ubiquinone oxidoreductase subunit AB1), acts as a powerful cardio-protector by enhancing mitochondrial energy biogenesis. In particular, NDUFAB1 coordinates the assembly of respiratory complexes I, II, and III and supercomplexes, conferring greater capacity and efficiency of mitochondrial energy metabolism.
    [Show full text]
  • A Comprehensive Review of Our Current Understanding of Red Blood Cell (RBC) Glycoproteins
    membranes Review A Comprehensive Review of Our Current Understanding of Red Blood Cell (RBC) Glycoproteins Takahiko Aoki Laboratory of Quality in Marine Products, Graduate School of Bioresources, Mie University, 1577 Kurima Machiya-cho, Mie, Tsu 514-8507, Japan; [email protected]; Tel.: +81-59-231-9569; Fax: +81-59-231-9557 Received: 18 August 2017; Accepted: 24 September 2017; Published: 29 September 2017 Abstract: Human red blood cells (RBC), which are the cells most commonly used in the study of biological membranes, have some glycoproteins in their cell membrane. These membrane proteins are band 3 and glycophorins A–D, and some substoichiometric glycoproteins (e.g., CD44, CD47, Lu, Kell, Duffy). The oligosaccharide that band 3 contains has one N-linked oligosaccharide, and glycophorins possess mostly O-linked oligosaccharides. The end of the O-linked oligosaccharide is linked to sialic acid. In humans, this sialic acid is N-acetylneuraminic acid (NeuAc). Another sialic acid, N-glycolylneuraminic acid (NeuGc) is present in red blood cells of non-human origin. While the biological function of band 3 is well known as an anion exchanger, it has been suggested that the oligosaccharide of band 3 does not affect the anion transport function. Although band 3 has been studied in detail, the physiological functions of glycophorins remain unclear. This review mainly describes the sialo-oligosaccharide structures of band 3 and glycophorins, followed by a discussion of the physiological functions that have been reported in the literature to date. Moreover, other glycoproteins in red blood cell membranes of non-human origin are described, and the physiological function of glycophorin in carp red blood cell membranes is discussed with respect to its bacteriostatic activity.
    [Show full text]
  • Time Dependent Pathway Activation of Signalling Cascades in Rat Organs After Short-Term Hyperoxia
    International Journal of Molecular Sciences Article Time Dependent Pathway Activation of Signalling Cascades in Rat Organs after Short-Term Hyperoxia Jochen Hinkelbein 1,*,†, Stefan Braunecker 2,† ID , Matthias Danz 3, Lennert Böhm 4 and Andreas Hohn 1 1 Department of Anesthesiology and Intensive Care Medicine, University Hospital of Cologne, 50937 Cologne, Germany; [email protected] 2 Department of Anaesthesiology, St George’s University Hospital, NHS Foundation Trust, London SW17 0QT, UK; [email protected] 3 Department of Anesthesiology, St. Martinus Hospital, 57462 Olpe, Germany; [email protected] 4 Emergency Department, University of Duesseldorf, 40225 Düsseldorf, Germany; [email protected] * Correspondence: [email protected]; Tel.: +49-221-478-82042 † These authors contributed equally to this work. Received: 1 May 2018; Accepted: 29 June 2018; Published: 4 July 2018 Abstract: Administration of oxygen is one of the most common interventions in medicine. Previous research showed that differential regulated proteins could be linked to hyperoxia-associated signaling cascades in different tissues. However, it still remains unclear which signaling pathways are activated by hyperoxia. The present study analyses hyperoxia-induced protein alterations in lung, brain, and kidney tissue using a proteomic and bioinformatic approach. Pooled data of 36 Wistar rats exposed to hyperoxia were used. To identify possible hyperoxia biomarkers, and to evaluate the relationship between protein alterations in hyperoxia affected organs and blood, proteomics data from brain, lung, and kidney were analyzed. Functional network analyses (IPA®, PathwaysStudio®, and GENEmania®) in combination with hierarchical cluster analysis (Perseus®) was used to identify relevant pathways and key proteins. Data of 54 2D-gels with more than 2500 significantly regulated spots per gel were collected.
    [Show full text]
  • GLUT-1 Content and Interaction with Stomatin in Red Blood Cells from Species Without Vitamin C Biosynthesis and Their Relevance for Diabetes Mellitus Type 1
    GLUT-1 content and interaction with stomatin in red blood cells from species without vitamin C biosynthesis and their relevance for diabetes mellitus type 1 Dissertation zur Erlangung eines Doktorgrades der Naturwissenschaften ( Dr. rer. nat. ) Fakultät Naturwissenschaften Universität Hohenheim Institut für biologische Chemie und Ernährungswissenschaften vorgelegt von Tabea Caroline Frey aus Freiburg im Breisgau 2016 Dekan: Prof. Dr. Heinz Breer Erstgutachter: Prof. Dr. Hans Konrad Biesalski Zweitgutachter: Prof. Dr. Jan Frank Eingereicht am: 22.04.2016 Mündliche Prüfung: 28.07.2016 Die vorliegende Arbeit wurde am 27.06.2016 von der Fakultät Naturwissenschaften der Universität Hohenheim als 'Dissertation zur Erlangung des Doktorgrades der Naturwissenschaften' angenommen. Abstract Ascorbic acid is commonly known as vitamin C. By denition, vitamins, with the exception of vitamin D, are substances which can not be synthesized but are essential for the organism. In this light vitamin C is special. It is hypothesized that millions of years ago some species, like primates, guinea pigs and fruit bats, lost the ability to synthesize ascorbate from glucose due to an inactivation of an enzyme called L-gulono-g-lactone oxidase. Since then, these species have been dependent on dietary intake of this micronutrient. Ascorbate is not only the most ecient watersoluble antioxidant, but also an important cofactor in neuro-transmitter or collagen biosynthesis. An inadequate intake of this vitamin leads to scurvy. In 2008, a French researcher documented that all species that lost the ability to synthesize ascorbic acid express a dierent facilitative glucose transporter isoform (GLUT) in their erythrocytes. The expressed GLUT-1 transports not only glucose but also dehydroascorbate which is the oxidized form of vitamin C.
    [Show full text]
  • Erythrocyte Glut1 Triggers Dehydroascorbic Acid Uptake in Mammals Unable to Synthesize Vitamin C
    Erythrocyte Glut1 Triggers Dehydroascorbic Acid Uptake in Mammals Unable to Synthesize Vitamin C Ame´ lie Montel-Hagen,1 Sandrina Kinet,1 Nicolas Manel,1,4 Ce´ dric Mongellaz,1 Rainer Prohaska,2 Jean-Luc Battini,1 Jean Delaunay,3 Marc Sitbon,1 and Naomi Taylor1,* 1Institut de Ge´ ne´ tique Mole´ culaire de Montpellier, CNRS, Universite´ Montpellier I and II, Montpellier, France 2Max F. Perutz Laboratories, Department of Medical Biochemistry, Medical University of Vienna, Vienna Biocenter, Vienna A-1030, Austria 3He´ matologie, Hoˆ pital de Biceˆ tre, APHP, INSERM U779, Faculte´ de Me´ decine Paris-Sud, Le Kremlin-Biceˆ tre, France 4Present address: Skirball Institute of Biomolecular Medicine, NYU School of Medicine, New York, NY 10016, USA. *Correspondence: [email protected] DOI 10.1016/j.cell.2008.01.042 SUMMARY Glut1 also transports L-dehydroascorbic acid (DHA), an oxi- dized intermediate of ascorbic acid (AA) (Bianchi and Rose, Of all cells, human erythrocytes express the highest 1986; Rumsey et al., 1997; Vera et al., 1993). DHA entry via Glut level of the Glut1 glucose transporter. However, the family transporters was initially investigated because of the struc- regulation and function of Glut1 during erythropoiesis tural similarities between DHA and glucose. Once transported into are not known. Here, we report that glucose transport the cell, DHA is immediately reduced to AA allowing a recycling actually decreases during human erythropoiesis de- of ascorbate (May, 1998). The uptake of AA into cells is mediated spite a >3-log increase in Glut1 transcripts. In con- by a distinct class of transporters; sodium-dependent vitamin C trast, Glut1-mediated transport of L-dehydroascorbic transporters (SVCT1 and SVCT2) whose expression profiles differ from those of Glut family members (Tsukaguchi et al., 1999).
    [Show full text]
  • Functional Characterization of Stomatin-Like Protein 2
    Functional characterization of stomatin-like protein 2 Panagiotis Mitsopoulos, M.Sc. Department of Microbiology and Immunology McGill University, Montréal, QC, Canada August 2017 A thesis submitted to McGill University in partial fulfillment of the requirements of the degree of Doctor of Philosophy © Panagiotis Mitsopoulos 2017 To my parents, Chris and Despina You provided me the opportunity to pursue anything Abstract Stomatin-like protein (SLP)-2 is a widely expressed and highly conserved protein identified in a proteome analysis of detergent-insoluble, glycolipid-enriched microdomains of human T cells activated through the antigen receptor. SLP-2 partitions mostly in mitochondria but a low abundance pool is present at the plasma membrane. SLP-2 is upregulated upon T cell activation, and its deletion in T cell-specific SLP-2 knockout mice results in a post-transcriptional defect in IL-2 production and decreased T cell responses. Mechanistically, SLP-2 binds the mitochondrial phospholipid cardiolipin and interacts with prohibitin (PHB) proteins, correlating with impaired cardiolipin compartmentalization and respiratory function in SLP-2-deficient T cells. In our current model of SLP-2 function, SLP-2 recruits PHBs to cardiolipin to form cardiolipin- enriched microdomains in the mitochondrial inner membrane. We hypothesize that these specialized microdomains may provide an environment necessary for the optimal assembly and function of multichain complexes, including respiratory supercomplexes and translation machinery. Respiratory chain complexes naturally associate to form supercomplexes for optimal electron transfer and function, and cardiolipin is known to be important in this process. We found that in the absence of SLP-2, T cells exhibited defective assembly of complex I-III2 and I- III2-IV1-3 supercomplexes but not of individual respiratory complexes.
    [Show full text]
  • Stomatin-Related Olfactory Protein, SRO, Specifically Expressed in The
    The Journal of Neuroscience, July 15, 2002, 22(14):5931–5937 Stomatin-Related Olfactory Protein, SRO, Specifically Expressed in the Murine Olfactory Sensory Neurons Ko Kobayakawa, Reiko Hayashi, Kenji Morita, Kazunari Miyamichi, Yuichiro Oka, Akio Tsuboi, and Hitoshi Sakano Department of Biophysics and Biochemistry, Graduate School of Science, University of Tokyo, Bunkyo-ku, Tokyo 113-0032, Japan We identified a stomatin-related olfactory protein (SRO) that is dant in most apical dendrites of OSNs, including olfactory cilia. specifically expressed in olfactory sensory neurons (OSNs). The Immunoprecipitation revealed that SRO associates with adeny- mouse sro gene encodes a polypeptide of 287 amino acids with lyl cyclase type III and caveolin-1 in the low-density membrane a calculated molecular weight of 32 kDa. SRO shares 82% fraction of olfactory cilia. Furthermore, anti-SRO antibodies sequence similarity with the murine stomatin, 78% with Cae- stimulated cAMP production in fractionated cilia membrane. norhabditis elegans MEC-2, and 77% with C. elegans UNC-1. SRO may play a crucial role in modulating odorant signals in the Unlike other stomatin-family genes, the sro transcript was lipid rafts of olfactory cilia. present only in OSNs of the main olfactory epithelium. No sro Key words: olfactory sensory neuron; olfactory cilia; stoma- expression was seen in vomeronasal neurons. SRO was abun- tin; MEC-2; lipid rafts; adenylyl cyclase; caveolin The vertebrate olfactory system can discriminate hundreds of To study the signaling and projection of OSNs, we prepared thousand of different odorants with odorant receptor (OR) mol- murine cDNA libraries from the OE and searched for genes that ecules expressed in the olfactory sensory neurons (OSNs) within are specifically expressed in the OE.
    [Show full text]
  • Stomatin Interacts with GLUT1/SLC2A1, Band 3/SLC4A1, and Aquaporin-1 in Human Erythrocyte Membrane Domains
    View metadata, citation and similar papers at core.ac.uk brought to you by CORE provided by Elsevier - Publisher Connector Biochimica et Biophysica Acta 1828 (2013) 956–966 Contents lists available at SciVerse ScienceDirect Biochimica et Biophysica Acta journal homepage: www.elsevier.com/locate/bbamem Stomatin interacts with GLUT1/SLC2A1, band 3/SLC4A1, and aquaporin-1 in human erythrocyte membrane domains Stefanie Rungaldier a, Walter Oberwagner a, Ulrich Salzer a, Edina Csaszar b, Rainer Prohaska a,⁎ a Max F. Perutz Laboratories (MFPL), Medical University of Vienna, Vienna, Austria b Mass Spectrometry Facility, MFPL, Vienna, Austria article info abstract Article history: The widely expressed, homo-oligomeric, lipid raft-associated, monotopic integral membrane protein Received 21 June 2012 stomatin and its homologues are known to interact with and modulate various ion channels and transporters. Received in revised form 20 October 2012 Stomatin is a major protein of the human erythrocyte membrane, where it associates with and modifies the Accepted 26 November 2012 glucose transporter GLUT1; however, previous attempts to purify hetero-oligomeric stomatin complexes for Available online 3 December 2012 biochemical analysis have failed. Because lateral interactions of membrane proteins may be short-lived and unstable, we have used in situ chemical cross-linking of erythrocyte membranes to fix the stomatin com- Keywords: fi fi Integral membrane proteins plexes for subsequent puri cation by immunoaf nity chromatography. To further enrich stomatin, we Lipid rafts prepared detergent-resistant membranes either before or after cross-linking. Mass spectrometry of the iso- Chemical cross-linking lated, high molecular, cross-linked stomatin complexes revealed the major interaction partners as glucose Protein–protein interaction transporter-1 (GLUT1), anion exchanger (band 3), and water channel (aquaporin-1).
    [Show full text]
  • The Endosome Is a Master Regulator of Plasma Membrane Collagen Fibril Assembly
    bioRxiv preprint doi: https://doi.org/10.1101/2021.03.25.436925; this version posted March 25, 2021. The copyright holder for this preprint (which was not certified by peer review) is the author/funder. All rights reserved. No reuse allowed without permission. The endosome is a master regulator of plasma membrane collagen fibril assembly 1Joan Chang*, 1Adam Pickard, 1Richa Garva, 1Yinhui Lu, 2Donald Gullberg and 1Karl E. Kadler* 1Wellcome Centre for Cell-Matrix Research, Faculty of Biology, Medical and Health, University of Manchester, Michael Smith Building, Oxford Road, Manchester M13 9PT UK, 2Department of Biomedicine and Center for Cancer Biomarkers, Norwegian Center of Excellence, University of Bergen, Norway. * Co-corresponding authors: JC email: [email protected] (orcid.org/0000-0002-7283- 9759); KEK email: [email protected] (orcid.org/0000-0003-4977-4683) Keywords: collagen-I, endocytosis, extracellular matrix, fibril, fibrillogenesis, integrin-a11, trafficking, VPS33b, [abstract] [149 word max] Collagen fibrils are the principal supporting elements in vertebrate tissues. They account for 25% of total protein mass, exhibit a broad range of size and organisation depending on tissue and stage of development, and can be under circadian clock control. Here we show that the remarkable dynamic pleomorphism of collagen fibrils is underpinned by a mechanism that distinguishes between collagen secretion and initiation of fibril assembly, at the plasma membrane. Collagen fibrillogenesis occurring at the plasma membrane requires vacuolar protein sorting (VPS) 33b (which is under circadian clock control), collagen-binding integrin-a11 subunit, and is reduced when endocytosis is inhibited. Fibroblasts lacking VPS33b secrete soluble collagen without assembling fibrils, whereas constitutive over-expression of VPS33b increases fibril number with loss of fibril rhythmicity.
    [Show full text]
  • Acute Regulation of Glut1 Function: the Role Of
    ACUTE REGULATION OF GLUT1 FUNCTION: THE ROLE OF DETERGENT-RESISTANT MEMBRANE DOMAINS by DARRELL RUBIN Submitted in partial fulfillment of the requirements For the degree of Doctor of Philosophy Thesis Adviser: Dr. Faramarz Ismail-Beigi Department of Pathology CASE WESTERN RESERVE UNIVERSITY August 2004 Copyright © 2004 by Darrell Casimir Rubin All rights reserved CASE WESTERN RESERVE UNIVERSITY SCHOOL OF GRADUATE STUDIES We hereby approve the dissertation of ______________________________________________________ candidate for the Ph.D. degree *. (signed)_______________________________________________ (chair of the committee) ________________________________________________ ________________________________________________ ________________________________________________ ________________________________________________ ________________________________________________ (date) _______________________ *We also certify that written approval has been obtained for any proprietary material contained therein. Dedication To my family and all the people providing inspiration or motivation along the way. Table of Contents Table of Contents …………………………………………………………………. v List of Tables and Figures ………………………………………………………... vii Acknowledgements ………………………………………………………………... ix List of Abbreviations ……………………………………………………………... x Abstract ……………………………………………………………………………. xi Chapter 1 Introduction Members of the Glucose Transporter Family …………………….. 1 Clone 9 cells and the Regulation of Glucose Transport ………….. 6 Protocols for Subcellular Fractionation …………………………..
    [Show full text]
  • Organization of the Respiratory Supercomplexes in Cells with Defective Complex III: Structural Features and Metabolic Consequences
    life Review Organization of the Respiratory Supercomplexes in Cells with Defective Complex III: Structural Features and Metabolic Consequences Michela Rugolo , Claudia Zanna and Anna Maria Ghelli * Department of Pharmacy and Biotechnology, University of Bologna, 40126 Bologna, Italy; [email protected] (M.R.); [email protected] (C.Z.) * Correspondence: [email protected] Abstract: The mitochondrial respiratory chain encompasses four oligomeric enzymatic complexes (complex I, II, III and IV) which, together with the redox carrier ubiquinone and cytochrome c, catalyze electron transport coupled to proton extrusion from the inner membrane. The protonmotive force is utilized by complex V for ATP synthesis in the process of oxidative phosphorylation. Respiratory complexes are known to coexist in the membrane as single functional entities and as supramolecular aggregates or supercomplexes (SCs). Understanding the assembly features of SCs has relevant biomedical implications because defects in a single protein can derange the overall SC organization and compromise the energetic function, causing severe mitochondrial disorders. Here we describe in detail the main types of SCs, all characterized by the presence of complex III. We show that the genetic alterations that hinder the assembly of Complex III, not just the activity, cause a rearrangement of the architecture of the SC that can help to preserve a minimal energetic function. Finally, the major Citation: Rugolo, M.; Zanna, C.; metabolic disturbances associated with severe SCs perturbation due to defective complex III are Ghelli, A.M. Organization of the discussed along with interventions that may circumvent these deficiencies. Respiratory Supercomplexes in Cells with Defective Complex III: Keywords: respiratory complexes; respiratory supercomplexes; oxidative stress; mitochondrial DNA; Structural Features and Metabolic MTCYB mutations; cytochrome b; complex III; mitochondrial diseases Consequences.
    [Show full text]