DOCSLIB.ORG

Open Weina Sun Doctoral Dissertation

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Open Weina Sun Doctoral Dissertation The Pennsylvania State University The Graduate School College of Agricultural Science HENIPAVIRUS ASSEMBLY AND BUDDING A Dissertation in Pathobiology by Weina Sun © 2015 Weina Sun Submitted in Partial Fulfillments of the Requirements for the Degree of Doctor of Philosophy May 2015 The dissertation of Weina Sun was reviewed and approved* by the following: Anthony P. Schmitt Associate Professor of Molecular Virology Dissertation Advisor Chair of the Committee Director of Pathobiology Graduate Program K. Sandeep Prabhu Professor of Immunology and Molecular Toxicology Pamela A. Hankey Giblin Professor of Immunology Immunology and Infectious Disease Undergraduate Advisor Robert F. Paulson Professor of Veterinary and Biomedical Science Craig E. Cameron Professor of Biochemistry and Molecular Biology Eberly Chair in Biochemistry and Molecular Biology *Signatures are on file in the Graduate School ii ABSTRACT The emergence of a group of paramyxoviruses called henipavirus has caused fatal illness such as severe vasculitis and encephalitis which resulted in high fatality rate in both humans and animals since 1990’s in south Asia and Australia. Like other paramyxoviruses, the essential viral protein that organizes the process of henipavirus assembly and budding is the matrix protein (M), where it functions to link together the viral glycoproteins and ribonucleoprotein complex (RNPs). Current studies have sought out to investigate the host factors that are involved in virus assembly and budding by interacting with M protein to facilitate M functions. In an effort to identify host factors that are important for henipavirus assembly and budding, proteomics-based approach was performed to identify host proteins that interact with viral M protein. Here, we affinity purified viral M proteins by FPLC and identified co-purifying host proteins by mass spectrometry. Co- immunoprecipitation was used as a secondary screening of protein candidates identified from mass spec results. One of the host proteins candidates, AP3B1, the beta subunit of AP-3 complex, was selected for further investigations. Importantly, a 29 amino acids polypeptide derived from AP3B1 hinge domain was identified as the minimal fragment to bind M protein as well as effectively inhibit henipavirus-like particles (VLPs) production. This inhibitory effect is due to disruption of M protein association with membrane. Additionally, in AP3B1 depleted cells by siRNA knockdown, Nipah VLP production is significantly reduced. By immunofluorescence iii microscopy, M protein colocalization with endogenous AP3B1 was also observed in mammalian cells. Our results suggested that AP-3 might play an important role in M protein functions. Henipaviruses have two types of surface glycoproteins, the attachment protein (G) and fusion protein (F). They were responsible for virus entry by mediating virus attachment to cell receptors and fusion with cell membrane. Numerous studies have illustrated how the virus entry occurs under the coordination of G and F proteins. However, little is known about how glycoproteins assemble into virions or VLPs. We know that M protein is the main driving force of particles formation, so we wondered how M protein coordinates with G protein and F protein to facilitate their trafficking and assembly into virions or VLPs. Here, under the unique pathway of HeV F protein identified by our collaborator Dr. Rebecca Dutch at University of Kentucky, we have found that HeV M protein, F protein and G protein all partially colocalized with Rab11a-REs, and overexpression of a DN Rab11a could significantly inhibit Hendra M-VLPs as well as F-VLPs formation in the cells. Interestingly, in cells that express inhibitory AP3B1 Hinge domain, M colocalization with Rab11-REs was disrupted. Unlike M protein or F protein, G proteins releases poorly into particles when expressed alone in cells. We observed that G incorporation into VLPs could be facilitated by co-expression of M protein but not F protein, likely through G-M interaction at the plasma membrane. Moreover, we have found that F protein incorporation into VLPs depended on its endocytic trafficking event regardless its cleavage status, as an endocytosis defective mutant F S490A that was retained on iv the cell surface was shown to be significantly defective on particles assembly. On the other hand, HeV F protein still well incorporated into VLPs when its cleavage was prevented by a cathepsin L inhibitor, E-64d. Therefore, we hypothesized that henipavirus M and F proteins have separate mechanisms for trafficking to Rab11-REs, with the M protein trafficking facilitated by its interaction with AP3B1. M and cathepsin-cleaved F proteins must then assemble together within these compartments prior to their delivery to the cell surface for particle budding and assemble with G protein through G-M interaction at the plasma membrane. v TABLE OF CONTENTS LIST OF FIGURES ................................................................................................... ix LIST OF TABLES ..................................................................................................... xi LIST OF ABBREVIATIONS ...................................................................................... xii CHAPTER 1 ............................................................................................................ 1 Introduction .......................................................................................................... 1 1.1 Paramyxovirus classification and significance ............................................... 1 1.2 Paramyxovirus genome and replication ........................................................ 6 1.3 Paramyxovirus matrix (M) protein and virus assembly ............................... 10 1.4 Virus-like particles production ..................................................................... 15 1.5 Trafficking of paramyxovirus glycoproteins ................................................. 17 1.6 Adaptor protein (AP) complexes ................................................................. 20 1.7 Adaptor protein 3 (AP-3) complex ............................................................... 23 1.8 AP-3 complex and viral trafficking ............................................................... 24 1.9 Rab11 GTPase in negative-strand RNA virus assembly ................................ 26 1.10 Identification of protein-protein interactions between virus and host ......... 29 1.11 Preview ........................................................................................................... 31 CHAPTER 2 .......................................................................................................... 34 Materials and Methods ....................................................................................... 34 Plasmids .................................................................................................................. 34 Henipavirus M protein affinity purification and mass spectrometry ..................... 37 Coimmunoprecipitation .......................................................................................... 38 Measurements of VLP production .......................................................................... 39 Membrane flotation assays to measure M protein membrane association .......... 41 RNA interference (RNAi) ......................................................................................... 41 Immunofluorescence microscopy ........................................................................... 42 Brefeldin A (BFA) treatment ................................................................................... 44 Cathepsin L inhibition VLP assay ............................................................................. 45 Phosphoinositide 3-kinase (PI3K) inhibition VLP assay .......................................... 46 Sedimentation gradient analysis of Hendra VLPs density ...................................... 46 vi Subcellular fractionation ......................................................................................... 47 CHAPTER 3 .......................................................................................................... 49 Matrix Proteins of Nipah and Hendra Viruses Interact with Beta Subunits of AP-3 Complexes .......................................................................................................... 49 3.1 Introduction ...................................................................................................... 49 3.2 Results ............................................................................................................... 51 Identification of henipavirus M-associating host proteins by affinity purification and mass spectrometry ...................................................................................... 51 AP3B1 interacts with Henipavirus M proteins via its hinge domain .................. 55 Overexpression of M-binding, AP3B1-derived polypeptides blocks production of henipavirus VLPs ................................................................................................. 61 Depletion of AP3B1 from cells impairs Nipah VLP production ........................... 66 Partial co-localization of Nipah virus M protein with endogenous AP3B1 in transfected cells .................................................................................................. 68 3.3 Discussion .........................................................................................................
Load more

Recommended publications
  • Guide for Common Viral Diseases of Animals in Louisiana
    Sampling and Testing Guide for Common Viral Diseases of Animals in Louisiana Please click on the species of interest: Cattle Deer and Small Ruminants The Louisiana Animal Swine Disease Diagnostic Horses Laboratory Dogs A service unit of the LSU School of Veterinary Medicine Adapted from Murphy, F.A., et al, Veterinary Virology, 3rd ed. Cats Academic Press, 1999. Compiled by Rob Poston Multi-species: Rabiesvirus DCN LADDL Guide for Common Viral Diseases v. B2 1 Cattle Please click on the principle system involvement Generalized viral diseases Respiratory viral diseases Enteric viral diseases Reproductive/neonatal viral diseases Viral infections affecting the skin Back to the Beginning DCN LADDL Guide for Common Viral Diseases v. B2 2 Deer and Small Ruminants Please click on the principle system involvement Generalized viral disease Respiratory viral disease Enteric viral diseases Reproductive/neonatal viral diseases Viral infections affecting the skin Back to the Beginning DCN LADDL Guide for Common Viral Diseases v. B2 3 Swine Please click on the principle system involvement Generalized viral diseases Respiratory viral diseases Enteric viral diseases Reproductive/neonatal viral diseases Viral infections affecting the skin Back to the Beginning DCN LADDL Guide for Common Viral Diseases v. B2 4 Horses Please click on the principle system involvement Generalized viral diseases Neurological viral diseases Respiratory viral diseases Enteric viral diseases Abortifacient/neonatal viral diseases Viral infections affecting the skin Back to the Beginning DCN LADDL Guide for Common Viral Diseases v. B2 5 Dogs Please click on the principle system involvement Generalized viral diseases Respiratory viral diseases Enteric viral diseases Reproductive/neonatal viral diseases Back to the Beginning DCN LADDL Guide for Common Viral Diseases v.
    [Show full text]
  • Arenaviridae Astroviridae Filoviridae Flaviviridae Hantaviridae
    Hantaviridae 0.7 Filoviridae 0.6 Picornaviridae 0.3 Wenling red spikefish hantavirus Rhinovirus C Ahab virus * Possum enterovirus * Aronnax virus * * Wenling minipizza batfish hantavirus Wenling filefish filovirus Norway rat hunnivirus * Wenling yellow goosefish hantavirus Starbuck virus * * Porcine teschovirus European mole nova virus Human Marburg marburgvirus Mosavirus Asturias virus * * * Tortoise picornavirus Egyptian fruit bat Marburg marburgvirus Banded bullfrog picornavirus * Spanish mole uluguru virus Human Sudan ebolavirus * Black spectacled toad picornavirus * Kilimanjaro virus * * * Crab-eating macaque reston ebolavirus Equine rhinitis A virus Imjin virus * Foot and mouth disease virus Dode virus * Angolan free-tailed bat bombali ebolavirus * * Human cosavirus E Seoul orthohantavirus Little free-tailed bat bombali ebolavirus * African bat icavirus A Tigray hantavirus Human Zaire ebolavirus * Saffold virus * Human choclo virus *Little collared fruit bat ebolavirus Peleg virus * Eastern red scorpionfish picornavirus * Reed vole hantavirus Human bundibugyo ebolavirus * * Isla vista hantavirus * Seal picornavirus Human Tai forest ebolavirus Chicken orivirus Paramyxoviridae 0.4 * Duck picornavirus Hepadnaviridae 0.4 Bildad virus Ned virus Tiger rockfish hepatitis B virus Western African lungfish picornavirus * Pacific spadenose shark paramyxovirus * European eel hepatitis B virus Bluegill picornavirus Nemo virus * Carp picornavirus * African cichlid hepatitis B virus Triplecross lizardfish paramyxovirus * * Fathead minnow picornavirus
    [Show full text]
  • Evidence to Support Safe Return to Clinical Practice by Oral Health Professionals in Canada During the COVID-19 Pandemic: a Repo
    Evidence to support safe return to clinical practice by oral health professionals in Canada during the COVID-19 pandemic: A report prepared for the Office of the Chief Dental Officer of Canada. November 2020 update This evidence synthesis was prepared for the Office of the Chief Dental Officer, based on a comprehensive review under contract by the following: Paul Allison, Faculty of Dentistry, McGill University Raphael Freitas de Souza, Faculty of Dentistry, McGill University Lilian Aboud, Faculty of Dentistry, McGill University Martin Morris, Library, McGill University November 30th, 2020 1 Contents Page Introduction 3 Project goal and specific objectives 3 Methods used to identify and include relevant literature 4 Report structure 5 Summary of update report 5 Report results a) Which patients are at greater risk of the consequences of COVID-19 and so 7 consideration should be given to delaying elective in-person oral health care? b) What are the signs and symptoms of COVID-19 that oral health professionals 9 should screen for prior to providing in-person health care? c) What evidence exists to support patient scheduling, waiting and other non- treatment management measures for in-person oral health care? 10 d) What evidence exists to support the use of various forms of personal protective equipment (PPE) while providing in-person oral health care? 13 e) What evidence exists to support the decontamination and re-use of PPE? 15 f) What evidence exists concerning the provision of aerosol-generating 16 procedures (AGP) as part of in-person
    [Show full text]
  • Accepted Version
    Article The cellular protein TIP47 restricts Respirovirus multiplication leading to decreased virus particle production BAMPI, Carole, et al. Abstract The cellular tail-interacting 47-kDa protein (TIP47) acts positively on HIV-1 and vaccinia virus production. We show here that TIP47, in contrast, acts as a restriction factor for Sendai virus production. This conclusion is supported by the occurrence of increased or decreased virus production upon its suppression or overexpression, respectively. Pulse-chase metabolic labeling of viral proteins under conditions of TIP47 suppression reveals an increased rate of viral protein synthesis followed by increased incorporation of viral proteins into virus particles. TIP47 is here described for the first time as a viral restriction factor that acts by limiting viral protein synthesis. Reference BAMPI, Carole, et al. The cellular protein TIP47 restricts Respirovirus multiplication leading to decreased virus particle production. Virus Research, 2013, vol. 173, no. 2, p. 354-63 DOI : 10.1016/j.virusres.2013.01.006 PMID : 23348195 Available at: http://archive-ouverte.unige.ch/unige:28890 Disclaimer: layout of this document may differ from the published version. 1 / 1 Virus Research 173 (2013) 354–363 Contents lists available at SciVerse ScienceDirect Virus Research journa l homepage: www.elsevier.com/locate/virusres The cellular protein TIP47 restricts Respirovirus multiplication leading to decreased virus particle production a a,b,1 c,d Carole Bampi , Anne-Sophie Gosselin Grenet , Grégory Caignard
    [Show full text]
  • Evolution and Diversity of Bat and Rodent Paramyxoviruses from North America Brendan B
    bioRxiv preprint doi: https://doi.org/10.1101/2021.07.01.450817; this version posted July 2, 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. Evolution and diversity of bat and rodent Paramyxoviruses from North America Brendan B. Larsen1, Sophie Gryseels1,2*, Hans W. Otto1, Michael Worobey1 1Department of Ecology and Evolutionary Biology, University of Arizona, Tucson, AZ 2 Department of Microbiology, Immunology and Transplantation, Rega Institute, KU Leuven, Laboratory of Clinical and Evolutionary Virology, Leuven, Belgium * Current addresses: Evolutionary Ecology group, Department Biology, University of Antwerp, Belgium OD Taxonomy and Phylogeny, Royal Belgian Institute of Natural Sciences, Belgium Abstract Paramyxoviruses are a diverse group of negative-sense, single-stranded RNA viruses of which several species cause significant mortality and morbidity. In recent years the collection of paramyxoviruses sequences detected in wild mammals has substantially grown, however little is known about paramyxovirus diversity in North American mammals. To better understand natural paramyxovirus diversity, host range, and host specificity, we sought to comprehensively characterize paramyxoviruses across a range of diverse co-occurring wild small mammals in Southern Arizona. We used highly degenerate primers to screen fecal and urine samples and obtained a total of 55 paramyxovirus sequences from 12 rodent species and 6 bat species. We also performed illumina RNA-seq and de novo assembly on 14 of the positive samples to recover a total of 5 near full-length viral genomes.
    [Show full text]
  • Imbroglios of Viral Taxonomy: Genetic Exchange and Failings of Phenetic Approaches Jeffrey G
    JOURNAL OF BACTERIOLOGY, Sept. 2002, p. 4891–4905 Vol. 184, No. 17 0021-9193/02/$04.00ϩ0 DOI: 10.1128/JB.184.17.4891–4905.2002 Copyright © 2002, American Society for Microbiology. All Rights Reserved. Imbroglios of Viral Taxonomy: Genetic Exchange and Failings of Phenetic Approaches Jeffrey G. Lawrence,* Graham F. Hatfull, and Roger W. Hendrix Pittsburgh Bacteriophage Institute and Department of Biological Sciences, University of Pittsburgh, Pittsburgh, Pennsylvania 15260 Received 13 March 2002/Accepted 23 April 2002 The practice of classifying organisms into hierarchical groups originated with Aristotle and was codified into nearly immutable biological law by Linnaeus. The heart of taxonomy is the biological species, which forms the foundation for higher levels of classification. Whereas species have long been established among sexual eukaryotes, achieving a meaningful species concept for prokaryotes has been an onerous task and has proven exceedingly difficult for describing viruses and bacteriophages. Moreover, the assembly of viral “species” into higher-order taxonomic groupings has been even more tenuous, since these groupings were based initially on limited numbers of morphological features and more recently on overall genomic similarities. The wealth of nucleotide sequence information that catalyzed a revolution in the taxonomy of free-living organisms necessitates a reevaluation of the concept of viral species, genera, families, and higher levels of classification. Just as microbiologists discarded dubious morphological traits in favor of more accurate molecular yardsticks of evolutionary change, virologists can gain new insight into viral evolution through the rigorous analyses afforded by the molecular phylogenetics of viral genes. For bacteriophages, such dissections of genomic sequences reveal fundamental flaws in the Linnaean paradigm that necessitate a new view of viral evolution, classification, and taxonomy.
    [Show full text]
  • Dsrna SIGNALING in INNATE IMMUNITY and VIRAL INHIBITION
    dsRNA SIGNALING IN INNATE IMMUNITY AND VIRAL INHIBITION by LENETTE LIN LU Submitted in partial fulfillment of the requirements for the degree of Doctor of Philosophy Thesis Advisor: Dr. Ganes C. Sen Medical Scientist Training Program / Department of Molecular Biology and Microbiology- Molecular Virology Program CASE WESTERN RESERVE UNIVERSITY January, 2009 CASE WESTERN RESERVE UNIVERSITY SCHOOL OF GRADUATE STUDIES We hereby approve the thesis/dissertation of _______________Lenette Lu_____________________________ candidate for the ____PhD_________________________degree *. (signed)___Robert Silverman____________________________ (chair of the committee) ____Jonathon Karn_______________________________ ____Ganes C Sen_________________________________ ____Clifford V Harding III_________________________ ____George R Stark_______________________________ ________________________________________________ (date) ____June 6 2008_______ We also certify that written approval has been obtained for any proprietary material contained therein. TABLE OF CONTENTS LIST OF TABLES……………………………………………………………. ………….7 LIST OF FIGURES…………………………………………………………... ………….8 ACKNOWLEDGEMENTS…………………………………………………….………..10 LIST OF ABBREVIATIONS…………………………………………………. ………..13 ABSTRACT………………………………………………………………………….…..16 CHAPTER 1. INTRODUCTION………………………………………………………..18 Innate and Adaptive Immunity…………………………………………………..18 Pathogen Associated Molecular Patterns and their Receptors in Innate Immunity……………………………………………………………….19 Receptors that respond to dsRNA………………………………………………..23 Extracellular
    [Show full text]
  • Pneumoviruses and Metapneumoviruses
    Pneumoviruses and Metapneumoviruses Edited by Helene F. Rosenberg & Joseph B. Domachowske Printed Edition of the Special Issue Published in Viruses www.mdpi.com/journal/viruses Helene F. Rosenberg and Joseph B. Domachowske (Eds.) Pneumoviruses and Metapneumoviruses This book is a reprint of the special issue that appeared in the online open access journal Viruses (ISSN 1999-4915) in 2013 (available at: http://www.mdpi.com/journal/viruses/special_issues/pneumoviruses_metapneumoviruses). Guest Editors Helene F. Rosenberg National Institute of Allergy and Infectious Diseases (NIAID) Bethesda, MD, USA Joseph B. Domachowske SUNY Upstate Medical University Syracuse, NY, USA Editorial Office MDPI AG Klybeckstrasse 64 Basel, Switzerland Publisher Shu-Kun Lin Production Editor Matthias Burkhalter 1. Edition 2013 MDPI • Basel • Beijing ISBN 978-3-03842-049-1 © 2013 by the authors; licensee MDPI, Basel, Switzerland. All articles in this volume are Open Access distributed under the Creative Commons Attribution 3.0 license (http://creativecommons.org/licenses/by/3.0/), which allows users to download, copy and build upon published articles even for commercial purposes, as long as the author and publisher are properly credited, which ensures maximum dissemination and a wider impact of our publications. However, the dissemination and distribution of copies of this book as a whole is restricted to MDPI, Basel, Switzerland. Table of Contents Preface ....................................................................................................................................................vii 1. General Review Lenneke E. M. Haas, Steven F. T. Thijsen, Leontine van Elden and Karen A. Heemstra Human Metapneumovirus in Adults......................................................................................................... 1 Reprinted from Viruses 2013, 5(1), 87-110; doi:10.3390/v501008 http://www.mdpi.com/1999-4915/5/1/87 2.
    [Show full text]
  • Prevention of Respiratory Syncytial Virus Attachment Protein Cleavage in Vero Cells Rescues Infectivity of Progeny Virions for Primary Human Airway Cultures
    Prevention of Respiratory Syncytial Virus Attachment Protein Cleavage in Vero Cells Rescues Infectivity of Progeny Virions for Primary Human Airway Cultures DISSERTATION Presented in Partial Fulfillment of the Requirements for the Degree Doctor of Philosophy in the Graduate School of The Ohio State University By Jacqueline D. Corry, B.A. Graduate Program in Integrated Biomedical Science Program The Ohio State University 2015 Dissertation Committee: Mark E. Peeples, Ph.D.—Advisor Douglas M. McCarty, Ph.D. Ian Davis, DVM, Ph.D. Stefan Niewiesk, DVM, Ph.D. Copyright by Jacqueline D. Corry 2015 Abstract Live attenuated respiratory syncytial virus (RSV) vaccine candidates are produced in Vero cells, a cell line that cleaves the attachment (G) glycoprotein. As a result, Vero- derived virus is 5-fold less infectious for primary well-differentiated human airway epithelial (HAE) cultures than virus grown in HeLa. HAE cultures are isolated directly from the human airways, so it is likely that Vero-grown vaccine virus would be similarly inefficient at initiating infection of the nasal epithelium following vaccination, requiring a larger inoculum, thereby raising the cost per dose. Using protease inhibitors with increasing specificity, we identified cathepsin L as the responsible protease and confirmed that virus grown in the presence of protease inhibitors was more infectious for HAE cultures. Our evidence suggests that the G protein interacts with cathepsin L in the late endosome or lysosome via endocytic recycling. While essential for Nipah virus F protein cleavage, endocytic recycling is detrimental to the production of infectious RSV from Vero cells. We found that cathepsin L is able to cleave the G protein in Vero-grown, but not in HeLa-grown virions suggesting a difference in G protein posttranslational modification.
    [Show full text]
  • The AFIP's Department of Veterinary Pathology & the American Registry of Pathology Dedicate This Pathology of Laboratory A
    The AFIP’s Department of Veterinary Pathology & the American Registry of Pathology dedicate this Pathology of Laboratory Animals syllabus to COL (Ret.) William Inskeep II. COL (Ret.) William Inskeep II 10 October 1949 – 2 July 2005 COL Inskeep entered active military service in 1971, with a Reserve Officer Training Corps commission in the US Air Force. He served five years as a Minuteman Missile Launch Officer at Francis E. Warren Air Force Base, Cheyenne, Wyoming. He was a graduate of Colorado State University and entered the US Army Veterinary Corps in June 1980. COL Inskeep was a Diplomate of the American College of Veterinary Pathologists and held the Surgeon General’s “A” Proficiency Designator in the field of Veterinary Pathology. He was also a member of the Order of Military Medical Merit. COL Inskeep had many diversified and challenging assignments as a Veterinary Corps Officer, his most recent being the Chair, Department of Veterinary Pathology, AFIP, 1997-2003. He also served as Veterinary Pathology Consultant to the Army Surgeon General, 2000-2003, as AFIP Deputy Director Army, 1998-2001, and as DoD Liaison Officer to the US Department of Agriculture, 1996-2000. COL Inskeep was a graduate of the Army War College. His military awards include Defense Superior Service Medal, Legion of Merit, Meritorious Service Medal with oak leaf cluster, Joint Service Commendation Medal, Army and Air Force Commendation Medals, and Army Staff Badge. COL Inskeep had many accomplishments over his 28 year military career. However, we will remember him most as a dedicated, enthusiastic mentor and teacher of many residents at the AFIP.
    [Show full text]
  • (12) United States Patent (10) Patent No.: US 8,883,481 B2 Dormitzer Et Al
    USOO8883481 B2 (12) United States Patent (10) Patent No.: US 8,883,481 B2 Dormitzer et al. (45) Date of Patent: Nov. 11, 2014 (54) REVERSE GENETICS METHODS FOR VIRUS (56) References Cited RESCUE U.S. PATENT DOCUMENTS (75) Inventors: Philipso Dormitzer, Weston, MA (US); 6,455,298 B1* 9/2002 Groner et al. .............. 435/235.1 Björn Keiner, Marburg (DE); Pirada 2008/O124803 A1 5/2008 Billeter et al. Suphaphiphat, Brookline, MA (US); Michael Franti, Quebec (CA); Peter FOREIGN PATENT DOCUMENTS Mason, Sommerville, MA (US); WO WO-2003/091401 4/2003 ennifer Uhlendorff, Neuss (DE): WO WO-2005/062820 7/2005 Mikhail Matrosovich, Marburg (DE) WO WO-2007/047459 7/2005 WO WO-2010/046335 4/2010 (73) Assignee: Novartis AG, Basel (CH) OTHER PUBLICATIONS (*) Notice: Subject to any disclaimer the term of this Wang et al. Cloning of the canine RNA polymerase I promoter and patent is extended or adjusted under 35 establishment of reverse genetics for influenza A and B in MDCK U.S.C. 154(b) by 17 days. cells. Virology Journal 2007, vol. 4, p. 102-113.* Nicolson et al. Generation of influenza vaccine viruses on Vero cells (21) Appl. No.: 13/503,353 by reverse genetics: an H5N1 candidate vaccine strain produced under a quality system. Vaccine 2005, vol. 23, pp. 2943-2952.* Koudstaal et al. (Apr. 28, 2009). "Suitability of PerC6 cells to gen (22) PCT Filed: Oct. 20, 2010 erate epidemic and pandemic influenza vaccine strains by reverse genetics.” Vaccine 27(19):2588-93. (86). PCT No.: PCT/B2010/054752 Neumann (2003).
    [Show full text]
  • Non-Canonical Translation in RNA Viruses Andrew E
    Journal of General Virology (2012), 93, 1385–1409 DOI 10.1099/vir.0.042499-0 Review Non-canonical translation in RNA viruses Andrew E. Firth and Ian Brierley Correspondence Division of Virology, Department of Pathology, University of Cambridge, Tennis Court Road, Andrew E. Firth Cambridge CB2 1QP, UK [email protected] Viral protein synthesis is completely dependent upon the translational machinery of the host cell. However, many RNA virus transcripts have marked structural differences from cellular mRNAs that preclude canonical translation initiation, such as the absence of a 59 cap structure or the presence of highly structured 59UTRs containing replication and/or packaging signals. Furthermore, whilst the great majority of cellular mRNAs are apparently monocistronic, RNA viruses must often express multiple proteins from their mRNAs. In addition, RNA viruses have very compact genomes and are under intense selective pressure to optimize usage of the available sequence space. Together, these features have driven the evolution of a plethora of non-canonical translational mechanisms in RNA viruses that help them to meet these challenges. Here, we review the mechanisms utilized by RNA viruses of eukaryotes, focusing on internal ribosome entry, leaky scanning, non-AUG initiation, ribosome shunting, reinitiation, ribosomal frameshifting and stop- codon readthrough. The review will highlight recently discovered examples of unusual translational strategies, besides revisiting some classical cases. Introduction events in translation of eukaryotic cellular mRNAs, the vast majority of which bear a 59 cap structure (m7G) and a 39 No virus encodes its own ribosome. Indeed, it has been poly(A) tail. Translation can be divided into four stages: proposed that the distinction between cellular life and the initiation, elongation, termination and ribosome recyc- virus world could be based simply on whether an entity en- ling.
    [Show full text]