DOCSLIB.ORG

Regulation of Vaccinia Virus Replication: a Story of Viral Mimicry and a Novel Antagonistic Relationship Between Vaccinia Kinase and Pseudokinase Annabel T

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Regulation of Vaccinia Virus Replication: a Story of Viral Mimicry and a Novel Antagonistic Relationship Between Vaccinia Kinase and Pseudokinase Annabel T University of Nebraska - Lincoln DigitalCommons@University of Nebraska - Lincoln Dissertations and Theses in Biological Sciences Biological Sciences, School of 4-2019 Regulation of Vaccinia Virus Replication: a Story of Viral Mimicry and a Novel Antagonistic Relationship Between Vaccinia Kinase and Pseudokinase Annabel T. Olson University of Nebraska-Lincoln, [email protected] Follow this and additional works at: https://digitalcommons.unl.edu/bioscidiss Part of the Biology Commons Olson, Annabel T., "Regulation of Vaccinia Virus Replication: a Story of Viral Mimicry and a Novel Antagonistic Relationship Between Vaccinia Kinase and Pseudokinase" (2019). Dissertations and Theses in Biological Sciences. 106. https://digitalcommons.unl.edu/bioscidiss/106 This Article is brought to you for free and open access by the Biological Sciences, School of at DigitalCommons@University of Nebraska - Lincoln. It has been accepted for inclusion in Dissertations and Theses in Biological Sciences by an authorized administrator of DigitalCommons@University of Nebraska - Lincoln. REGULATION OF VACCINIA VIRUS REPLICATION: A STORY OF VIRAL MIMICRY AND A NOVEL ANTAGONISTIC RELATIONSHIP BETWEEN VACCINIA KINASE AND PSEUDOKINASE by Annabel T. Olson A DISSERTATION Presented to the Faculty of The Graduate College at the University of Nebraska In Partial Fulfillment of Requirements For the Degree of Doctor of Philosophy Major: Biological Sciences (Genetics, Cell, and Molecular Biology) Under the Supervision of Professor Matthew S. Wiebe Lincoln, Nebraska April, 2019 SUPERVISORY COMMITTEE SIGNATURES SUPERVISORY COMMITTEE: Matt Wiebe, Ph.D. (Primary Advisor) Deb Brown, Ph.D. Audrey Atkin, Ph.D. Luwen Zhang, Ph.D. Asit Pattnaik, Ph.D. APPROVED Regulation of vaccinia virus replication: A story of viral mimicry and a novel antagonistic relationship between vaccinia kinase and pseudokinase Annabel T. Olson, Ph.D. University of Nebraska, 2019 Advisor: Matthew S. Wiebe Poxviruses employ sophisticated signaling pathways that thwart cellular defense mechanisms and simultaneously ensure viral factors are modulated properly. Yet, our understanding of these complex signaling networks are incomplete. For example, the vaccinia B1 kinase plays a vital role in inactivating the cellular antiviral factor BAF, and is suggested to orchestrate other pathways. B1 is highly conserved among poxviruses and exhibits a remarkable degree of similarity to VRKs, a family of cellular kinases, suggesting that the viral enzyme has evolved to mimic VRK activity. Indeed, B1 and VRKs have been demonstrated to target a shared substrate, the DNA binding protein BAF, elucidating a signaling pathway important for mitosis and the antiviral response. Our research further characterized the role of B1 during vaccinia infection to gain novel insights into its regulation and integration with cellular signaling pathways. We began by constructing and characterizing the first B1 deletion virus (ΔB1). Then using this virus, we tested the hypothesis that cellular VRKs can complement B1 function, and discovered a VRK2 role in facilitating DNA replication in the absence of B1. Study of the VRK2 mechanism revealed that B1 and VRK2 mediate DNA replication via an additional pathway that is BAF independent. We also utilized the ΔB1 virus in an experimental evolution assay to perform an unbiased search for suppressor mutations and identify novel pathways involving B1. Interestingly, our characterization of the adapted viruses reveals that mutations correlating with a loss of function of the vaccinia B12 pseudokinase provide a striking fitness enhancement to this virus. Next, B12 characterization showed a nuclear localization, unique for poxvirus proteins, that is related to its repressive function. Our data indicate that B12 is not a global repressor, but inhibits vaccinia replication in the absence of the B1 kinase. The mechanism of B12 partially depends on suppression of BAF antiviral activity. However, the parallel B12 pathway to restrict virus replication is less clear. Together, our studies of B1 and B12 present novel evidence that a paralogous kinase-pseudokinase pair can exhibit a unique epistatic relationship in a virus, and orchestrate yet-to-be-discovered nuclear events during infection. DEDICATION This doctoral dissertation thesis is dedicated to my parents David and Diana Olson who support me constantly through their love and prayers. By the graces of God my mom is with us today to see me graduate. My mom was diagnosed with stage IV pancreatic cancer in November 2015 and is now in remission since November 2018. She is truly our miracle. God is good. ACKNOWLEDGMENTS When they say it takes a village to raise a child, the same can be said for a doctoral recipient. This achievement was not obtained without the support and guidance of many mentors, colleagues/friends and family members. I feel extremely fortunate to have been advised by Matt Wiebe over my graduate education. His guidance in my personal life, professional development and training as a researcher has greatly contributed to my success as a virologist and growth as an individual. In addition to my training in virology research, learning from Matt’s leadership roles in our lab and at the faculty and university levels has been an outstanding example of what a good mentor and colleague looks like. From his example, I will strive to emulate his kindness and insistent drive in order to be a good mentor and colleague to others. Recognition of my graduate committee members Deb Brown, Audrey Atkin, Asit Pattnaik, and Luwen Zhang is also a necessity for their contributions to my development as a researcher. They challenged my critical thinking skills and ability to articulate my research achievements and proposals. Deb recognized my talent in soft skills and encouraged me to develop this area. She also provided an excellent example of a passionate and skilled educator and researchers who knows the importance of lab community. Each of my committee members welcomed informal meetings and these discussions were some of the best conversations, as I maneuvered through the challenges during my graduate degree. The previous and current members of the Wiebe lab have been extremely supportive over my time here. I am grateful for the training and patience of Augusta Jamin and April Wicklund during my first years of research in the Wiebe lab and the fellowship of Zhigang Wang, Angela Mercurio, Shawna Fitzwater, Amber Rico, Allie Linville, and Luisa Valencia. I am grateful for the guidance and friendship of my colleagues Jessica Hargarten, Yen Ahmad Hussin, Kay Kimpston-Burkgren, Nacho Correas, Dane Bowder, Alex Vogel, Anna Lampe, Nicole Bacheller, Eric Noel, Sara Privatt, and the graduate students at the NCV. The staff members at the NCV have been wonderful to work with, especially Jan Edwards who always greeted me with a warm smile and words of encouragement. My friends outside of graduate school have also been extremely supportive during this time in graduate school including Grace Regan Reid, Tim Reid, Kelsey Nelson, Allie Klug Dunham, Alicia Wagner Gotto, Katie Johnson Harmelink, Kairsten Nelson, Christie Gleason Lewandoski, and Connor Johnson. Lastly, my family has been my strong pillar throughout graduate school. I am so thankful for their many visits, long phone conversations and many prayers. The pilgrimages we took together and discussions of God have led me down this current path. Thank you Dad, Mom, Chris, Hailey, Logan, Sophia, Marty, Kathy, and Cordelia for bringing so much happiness and light into my life. Thank you Lord for all of these people and the ones that are not mentioned, but still impacted by successful achievement of becoming a doctoral recipient. PREFACE Chapters 3 and 4, expect section 4.1, have been published in Journal of Virology. (Olson, AT. Rico, AB. Wang, Z. Delhon, G. Wiebe, MS. Deletion of the Vaccinia B1 Kinase Reveals Essential Function of this Enzyme Complemented Partly by the Homologous Cellular Kinase VRK2. J Virol. 2017 Jul 12; 91(15). pii: e00635-17. doi: 10.1128/JVI.00635-17) Chapters 5 and 6, except parts of 5.3, 6.2, and 6.4-6.7, have been published in PLOS Pathogens. (Olson, AT. Wang, Z. Rico, AB. Wiebe, MS. A poxvirus pseudokinase represses viral DNA replication via a pathway antagonized by its paralog kinase. PLoS Pathog. 2019 Feb 15;15(2):e1007608. doi: 10.1371/journal.ppat.1007608) TABLE OF CONTENTS TITLE…………………………………………………………………………………......i SUMPERVISORY COMMITTEE SIGNATURES……………………………………ii ABSTRACT…………………………………………………………………………......iii DEDICATION…………………………………………………………………………...v ACKNOWLEGEMENTS………………………………………………………………vi PREFACE………………………………………………………………………………vii ABBREVIATIONS LIST……………………………………………………………...xiii FIGURES LIST…………………………………………………………………………xv CHAPTER 1 INTRODUCTION…………………………………………………………………….....1 1.1. POXVIRIDAE FAMILY…………………………………………………………...1 1.1.A. History…………………………………………………………..............................2 1.1.B. Genome………………………………………………………….............................3 1.1.C. Vaccinia virus life cycle…………………………………………………………....6 1.1.D. Innate immunity………………………………………………………………….10 1.1.E. Viral mimicry elicited for counteracting innate immunity………………………..14 1.2. VACCINIA B1 KINASE VIRUS MIMICRY OF HOST VACCINIA-RELATED KINASES…………………………………………………………..........................15 1.2.A. Vaccinia virus B1 kinase…………………………………………………………17 1.2.B. Cellular vaccinia-related kinases (VRKs) ……………………………………….19 1.2.C. Cellular barrier-to-autointegration factor (BAF) substrate……………………….21 1.3. ENZYME / PSEUDOENZYME
Load more

Recommended publications
  • United States Patent (19) 11 Patent Number: 5,766,598 Paoletti Et Al
    USOO5766598A United States Patent (19) 11 Patent Number: 5,766,598 Paoletti et al. 45) Date of Patent: Jun. 16, 1998 54 RECOMBINANTATTENUATED ALVAC Murphy. F., 1996. "Virus Taxonomy", in Fields Virology, CANARYPOXVIRUS EXPRESSION Third Edition, Fields et al., eds. Lippincott-Raven Publish WECTORS CONTAINING HETEROLOGOUS ers. Philadelphia, pp. 15-57. DNA SEGMENTS ENCODNG LENTWRAL Taylor et al., 1991. Vaccine 9(3):190-193. GENE PRODUCTS Berman et al., 1990, Nature 345:622-625. Girard et al., 1991 Proc. Natl. Acad. Sci., USA 88:542-546. 75 Inventors: Enzo Paoletti, Delmar; James Tartaglia. Schenectady; William Irvin Hu et al., 1986. Nature 320:537-540. Cox. Troy, all of N.Y. Ho et al., 1990, Eur, J. Immunol. 20:477-483. Virology, vol. 173, No. 1, issued Nov. 1989, S. Dallo et al. 73) Assignee: Wirogenetics Corporation, Troy, N.Y. "Humoral Immune Response Elicited by Highly Attenuated Variants of Vaccinia Virus and by an Attenuated Recombi nant Expressing HIV-1 Envelope Protein". pp. 323-328 21 Appl. No.: 303,275 (entire document). (22 Filed: Sep. 7, 1994 Virology, vol. 152, issued 1986. M.E. Perkus et al., “Inser tion and Deletion Mutants of Vaccinia Virus". pp. 285-297 Related U.S. Application Data (entire document). Nature, vol. 317, issued 31 Oct. 1985, R.M.L. Buller et al., 63 Continuation of Ser. No. 897,382, Jun. 11, 1992, abandoned, which is a continuation-in-part of Ser. No. 715,921, Jun. 14, "Decreased Virulence of Recombinant Vaccinia Virus 1991, abandoned, and Ser. No. 847,951, Mar. 6, 1992, which Expression Vectors is Associated With a Thymidine is a continuation-in-part of Ser.
    [Show full text]
  • Herpes Simplex Virus Type 1 Oril Is Not Required for Virus Infection In
    JOURNAL OF VIROLOGY, Nov. 1987, p. 3528-3535 Vol. 61, No. 11 0022-538X/87/113528-08$02.00/0 Copyright C 1987, American Society for Microbiology Herpes Simplex Virus Type 1 oriL Is Not Required for Virus Replication or for the Establishment and Reactivation of Latent Infection in Mice MARYELLEN POLVINO-BODNAR, PAULO K. ORBERG, AND PRISCILLA A. SCHAFFER* Laboratory of Tumor Virus Genetics, Dana-Farber Cancer Institute, and Department of Microbiology and Molecular Genetics, Harvard Medical School, Boston, Massachusetts 02115 Received 11 May 1987/Accepted 31 July 1987 During the course of experiments designed to isolate deletion mutants of herpes simplex virus type 1 in the gene encoding the major DNA-binding protein, ICP8, a mutant, d61, that grew efficiently in ICP8-expressing Vero cells but not in normal Vero cells was isolated (P. K. Orberg and P. A. Schaffer, J. Virol. 61:1136-1146, 1987). d61 was derived by cotransfection of ICP8-expressing Vero cells with infectious wild-type viral DNA and a plasmid, pDX, that contains an engineered 780-base-pair (bp) deletion in the ICP8 gene, as well as a spontaneous -55-bp deletion in OriL. Gel electrophoresis and Southern blot analysis indicated that d61 DNA carried both deletions present in pDX. The ability of d61 to replicate despite the deletion in OriL suggested that a functional OriL is not essential for virus replication in vitro. Because d61 harbored two mutations, a second mutant, ts+7, with a deletion in oriL-associated sequences and an intact ICP8 gene was constructed. Both d61 and ts+7 replicated efficiently in their respective permissive host cells, although their yields were slightly lower than those of control viruses with intact oriL sequences.
    [Show full text]
  • Topological Analysis of the Gp41 MPER on Lipid Bilayers Relevant to the Metastable HIV-1 Envelope Prefusion State
    Topological analysis of the gp41 MPER on lipid bilayers relevant to the metastable HIV-1 envelope prefusion state Yi Wanga,b, Pavanjeet Kaurc,d, Zhen-Yu J. Sune,1, Mostafa A. Elbahnasawya,b,2, Zahra Hayatic,d, Zhi-Song Qiaoa,b,3, Nhat N. Buic, Camila Chilea,b,4, Mahmoud L. Nasre,5, Gerhard Wagnere, Jia-Huai Wanga,f, Likai Songc, Ellis L. Reinherza,b,6, and Mikyung Kima,g,6 aLaboratory of Immunobiology, Department of Medical Oncology, Dana-Farber Cancer Institute, Boston, MA 02115; bDepartment of Medicine, Harvard Medical School, Boston, MA 02115; cNational High Magnetic Field Laboratory, Florida State University, Tallahassee, FL 32306; dDepartment of Physics, Florida State University, Tallahassee, FL 32306; eDepartment of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, Boston, MA 02115; fDepartment of Cancer Biology, Dana-Farber Cancer Institute, Boston, MA 02215; and gDepartment of Dermatology, Harvard Medical School, Boston, MA 02215 Edited by Peter Palese, Icahn School of Medicine at Mount Sinai, New York, NY, and approved September 23, 2019 (received for review July 18, 2019) The membrane proximal external region (MPER) of HIV-1 envelope immunologically vulnerable epitopes targeted by several of the most glycoprotein (gp) 41 is an attractive vaccine target for elicitation of broadly neutralizing antibodies (bNAbs) developed during the broadly neutralizing antibodies (bNAbs) by vaccination. However, course of natural HIV-1 infection (10–13). Insertion, deletion, current details regarding the quaternary structural organization of and mutations of residues in the MPER defined the functional the MPER within the native prefusion trimer [(gp120/41)3] are elu- importance of the MPER in Env incorporation, viral fusion, and sive and even contradictory, hindering rational MPER immunogen infectivity (14–16).
    [Show full text]
  • Viroporins: Structures and Functions Beyond Cell Membrane Permeabilization
    Editorial Viroporins: Structures and Functions beyond Cell Membrane Permeabilization José Luis Nieva 1,* and Luis Carrasco 2,* Received: 17 September 2015 ; Accepted: 21 September 2015 ; Published: 29 September 2015 Academic Editor: Eric O. Freed 1 Biophysics Unit (CSIC, UPV/EHU) and Biochemistry and Molecular Biology Department, University of the Basque Country (UPV/EHU), P.O. Box 644, 48080 Bilbao, Spain 2 Centro de Biología Molecular Severo Ochoa (CSIC, UAM), c/Nicolás Cabrera, 1, Universidad Autónoma de Madrid, Cantoblanco, 28049 Madrid, Spain * Correspondence: [email protected] (J.L.N.); [email protected] (L.C.); Tel.: +34-94-601-3353 (J.L.N.); +34-91-497-8450 (L.C.) Viroporins represent an interesting group of viral proteins that exhibit two sets of functions. First, they participate in several viral processes that are necessary for efficient production of virus progeny. Besides, viroporins interfere with a number of cellular functions, thus contributing to viral cytopathogenicity. Twenty years have elapsed from the first review on viroporins [1]; since then several reviews have covered the advances on viroporin structure and functioning [2–8]. This Special Issue updates and revises new emerging roles of viroporins, highlighting their potential use as antiviral targets and in vaccine development. Viroporin structure. Viroporins are usually short proteins with at least one hydrophobic amphipathic helix. Homo-oligomerization is achieved by helix–helix interactions in membranes rendering higher order structures, forming aqueous pores. Progress in viroporin structures during the last 2–3 years has in some instances provided a detailed knowledge of their functional architecture, including the fine definition of binding sites for effective inhibitors.
    [Show full text]
  • How Influenza Virus Uses Host Cell Pathways During Uncoating
    cells Review How Influenza Virus Uses Host Cell Pathways during Uncoating Etori Aguiar Moreira 1 , Yohei Yamauchi 2 and Patrick Matthias 1,3,* 1 Friedrich Miescher Institute for Biomedical Research, 4058 Basel, Switzerland; [email protected] 2 Faculty of Life Sciences, School of Cellular and Molecular Medicine, University of Bristol, Bristol BS8 1TD, UK; [email protected] 3 Faculty of Sciences, University of Basel, 4031 Basel, Switzerland * Correspondence: [email protected] Abstract: Influenza is a zoonotic respiratory disease of major public health interest due to its pan- demic potential, and a threat to animals and the human population. The influenza A virus genome consists of eight single-stranded RNA segments sequestered within a protein capsid and a lipid bilayer envelope. During host cell entry, cellular cues contribute to viral conformational changes that promote critical events such as fusion with late endosomes, capsid uncoating and viral genome release into the cytosol. In this focused review, we concisely describe the virus infection cycle and highlight the recent findings of host cell pathways and cytosolic proteins that assist influenza uncoating during host cell entry. Keywords: influenza; capsid uncoating; HDAC6; ubiquitin; EPS8; TNPO1; pandemic; M1; virus– host interaction Citation: Moreira, E.A.; Yamauchi, Y.; Matthias, P. How Influenza Virus Uses Host Cell Pathways during 1. Introduction Uncoating. Cells 2021, 10, 1722. Viruses are microscopic parasites that, unable to self-replicate, subvert a host cell https://doi.org/10.3390/ for their replication and propagation. Despite their apparent simplicity, they can cause cells10071722 severe diseases and even pose pandemic threats [1–3].
    [Show full text]
  • Assembly Lecture 11 Biology W3310/4310 Virology Spring 2014
    Assembly Lecture 11 Biology W3310/4310 Virology Spring 2014 “Anatomy is des.ny.” --SIGMUND FREUD All virions complete a common set of assembly reac3ons * common to all viruses common to many viruses ©Principles of Virology, ASM Press The structure of a virus parcle determines how it is formed ©Principles of Virology, ASM Press Assembly is dependent on host cell machinery • Cellular chaperones • Transport systems • Secretory pathway • Nuclear import and export machinery Concentrang components for assembly: Nothing happens fast in dilute solu1ons • Viral components oSen visible by light microscopy (‘factories’ or ‘inclusions’) • Concentrate proteins on internal membranes (poliovirus) • Negri bodies (rabies virus) Viral proteins have ‘addresses’ built into their structure • Membrane targeYng: Signal sequences, fa\y acid modificaons • Membrane retenYon signals • Nuclear localizaYon sequences (NLS) • Nuclear export signals 414 Localiza3on of viral proteins to the nucleus CHAPTER 12 Golgi apparatus Ribosome Rough endoplasmic reticulum Plasma membrane Py(VP1) + VP2/3 Ad hexon + 5 100 kDa Nuclear envelope: Outer nuclear membrane Inner nuclear membrane Nucleus Nuclear pore complex Mitochondrion Cytoskeleton: Influenza virus NP Intermediate filament Microtubule Actin filament bundle Extracellular matrix ©Principles of Virology, ASM Press Figure 12.1 Localization of viral proteins to the nucleus. The nucleus and major membrane-bound compartments of the cytoplasm, as well as components of the cytoskeleton, are illustrated schematically and not to scale. Viral proteins destined for the nucleus are synthesized by cytoplasmic polyribosomes, as illustrated for the infl uenza virus NP protein. They engage with the cytoplasmic face of the nuclear pore complex and are translocated into the nucleus by the protein import machinery of the host cell.
    [Show full text]
  • Transcriptomic Profiles of High and Low Antibody Responders to Smallpox
    Genes and Immunity (2013) 14, 277–285 & 2013 Macmillan Publishers Limited All rights reserved 1466-4879/13 www.nature.com/gene ORIGINAL ARTICLE Transcriptomic profiles of high and low antibody responders to smallpox vaccine RB Kennedy1,2, AL Oberg1,3, IG Ovsyannikova1,2, IH Haralambieva1,2, D Grill1,3 and GA Poland1,2 Despite its eradication over 30 years ago, smallpox (as well as other orthopox viruses) remains a pathogen of interest both in terms of biodefense and for its use as a vector for vaccines and immunotherapies. Here we describe the application of mRNA-Seq transcriptome profiling to understanding immune responses in smallpox vaccine recipients. Contrary to other studies examining gene expression in virally infected cell lines, we utilized a mixed population of peripheral blood mononuclear cells in order to capture the essential intercellular interactions that occur in vivo, and would otherwise be lost, using single cell lines or isolated primary cell subsets. In this mixed cell population we were able to detect expression of all annotated vaccinia genes. On the host side, a number of genes encoding cytokines, chemokines, complement factors and intracellular signaling molecules were downregulated upon viral infection, whereas genes encoding histone proteins and the interferon response were upregulated. We also identified a small number of genes that exhibited significantly different expression profiles in subjects with robust humoral immunity compared with those with weaker humoral responses. Our results provide evidence that differential gene regulation patterns may be at work in individuals with robust humoral immunity compared with those with weaker humoral immune responses. Genes and Immunity (2013) 14, 277–285; doi:10.1038/gene.2013.14; published online 18 April 2013 Keywords: Next-generation sequencing; mRNA-Seq; vaccinia virus; smallpox vaccine INTRODUCTION these 44 subjects had two samples (uninfected and vaccinia Vaccinia virus (VACV) is the immunologically cross-protective infected).
    [Show full text]
  • Ube1a Suppresses Herpes Simplex Virus-1 Replication
    viruses Article UBE1a Suppresses Herpes Simplex Virus-1 Replication Marina Ikeda 1 , Akihiro Ito 2, Yuichi Sekine 1 and Masahiro Fujimuro 1,* 1 Department of Cell Biology, Kyoto Pharmaceutical University, Kyoto 607-8412, Japan; [email protected] (M.I.); [email protected] (Y.S.) 2 Laboratory of Cell Signaling, School of Life Sciences, Tokyo University of Pharmacy and Life Sciences, Tokyo 192-0392, Japan; [email protected] * Correspondence: [email protected]; Tel.: +81-75-595-4717 Academic Editors: Magdalena Weidner-Glunde and Andrea Lipi´nska Received: 7 November 2020; Accepted: 1 December 2020; Published: 4 December 2020 Abstract: Herpes simplex virus-1 (HSV-1) is the causative agent of cold sores, keratitis, meningitis, and encephalitis. HSV-1-encoded ICP5, the major capsid protein, is essential for capsid assembly during viral replication. Ubiquitination is a post-translational modification that plays a critical role in the regulation of cellular events such as proteasomal degradation, protein trafficking, and the antiviral response and viral events such as the establishment of infection and viral replication. Ub-activating enzyme (E1, also named UBE1) is involved in the first step in the ubiquitination. However, it is still unknown whether UBE1 contributes to viral infection or the cellular antiviral response. Here, we found that UBE1a suppressed HSV-1 replication and contributed to the antiviral response. The UBE1a inhibitor PYR-41 increased HSV-1 production. Immunofluorescence analysis revealed that UBE1a highly expressing cells presented low ICP5 expression, and vice versa. UBE1a inhibition by PYR-41 and shRNA increased ICP5 expression in HSV-1-infected cells.
    [Show full text]
  • HSV-1 Single-Cell Analysis Reveals the Activation of Anti-Viral And
    RESEARCH ARTICLE HSV-1 single-cell analysis reveals the activation of anti-viral and developmental programs in distinct sub-populations Nir Drayman1,2*, Parthiv Patel1,2, Luke Vistain1,2, Savas¸Tay1,2* 1Institute for Molecular Engineering, The University of Chicago, Chicago, United States; 2Institute for Genomics and Systems Biology, The University of Chicago, Chicago, United States Abstract Viral infection is usually studied at the population level by averaging over millions of cells. However, infection at the single-cell level is highly heterogeneous, with most infected cells giving rise to no or few viral progeny while some cells produce thousands. Analysis of Herpes Simplex virus 1 (HSV-1) infection by population-averaged measurements has taught us a lot about the course of viral infection, but has also produced contradictory results, such as the concurrent activation and inhibition of type I interferon signaling during infection. Here, we combine live-cell imaging and single-cell RNA sequencing to characterize viral and host transcriptional heterogeneity during HSV-1 infection of primary human cells. We find extreme variability in the level of viral gene expression among individually infected cells and show that these cells cluster into transcriptionally distinct sub-populations. We find that anti-viral signaling is initiated in a rare group of abortively infected cells, while highly infected cells undergo cellular reprogramming to an embryonic-like transcriptional state. This reprogramming involves the recruitment of b-catenin to the host nucleus and viral replication compartments, and is required for late viral gene expression and progeny production. These findings uncover the transcriptional differences in cells with variable infection outcomes and shed new light on the manipulation of host pathways by HSV-1.
    [Show full text]
  • PF a Colonne V 5 2020 12 16.Xlsx
    Istanza per il riconoscimento dell'esenzione della Tassa Automobilistica Ex Art 7 Comma 4 della Legge Regionale 12 Maggio 2020 n. 9 Elenco delle richieste di rimborso Persone Fisiche Le istanze sono riportate nell'elenco ordinate in modo crescente. Il motivo dell'eventuale rifiuto è solo perché non è stato trovato un versamento corrispondente al veicolo indicato per l'annualità 2020.
    [Show full text]
  • HIV-1) CD4 Receptor and Its Central Role in Promotion of HIV-1 Infection
    MICROBIOLOGICAL REVIEWS, Mar. 1995, p. 63–93 Vol. 59, No. 1 0146-0749/95/$04.0010 Copyright q 1995, American Society for Microbiology The Human Immunodeficiency Virus Type 1 (HIV-1) CD4 Receptor and Its Central Role in Promotion of HIV-1 Infection STEPHANE BOUR,* ROMAS GELEZIUNAS,† AND MARK A. WAINBERG* McGill AIDS Centre, Lady Davis Institute-Jewish General Hospital, and Departments of Microbiology and Medicine, McGill University, Montreal, Quebec, Canada H3T 1E2 INTRODUCTION .........................................................................................................................................................63 RETROVIRAL RECEPTORS .....................................................................................................................................64 Receptors for Animal Retroviruses ........................................................................................................................64 CD4 Is the Major Receptor for HIV-1 Infection..................................................................................................65 ROLE OF THE CD4 CORECEPTOR IN T-CELL ACTIVATION........................................................................65 Structural Features of the CD4 Coreceptor..........................................................................................................65 Interactions of CD4 with Class II MHC Determinants ......................................................................................66 CD4–T-Cell Receptor Interactions during T-Cell Activation
    [Show full text]
  • HIV-1 Env Gp41 Transmembrane Domain Dynamics Are Modulated by Lipid, Wa- Ter, and Ion Interactions
    bioRxiv preprint doi: https://doi.org/10.1101/292326; this version posted March 30, 2018. 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 4.0 International license. HIV-1 Env gp41 Transmembrane Domain Dynamics are Modulated by Lipid, Wa- ter, and Ion Interactions L.R. Hollingsworth IV, J.A. Lemkul, D.R. Bevan, and A.M. Brown Running Title: gp41 trimer dynamics in a lipid membrane Keywords: molecular dynamics simulations, protein-membrane interactions, HIV-1 gp41, envelope glycoprotein 1 bioRxiv preprint doi: https://doi.org/10.1101/292326; this version posted March 30, 2018. 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 4.0 International license. Abstract The gp41 transmembrane domain (TMD) of the envelope glycoprotein (Env) of the human immunodeficiency virus (HIV) modulates the conformation of the viral enve- lope spike, the only druggable target on the surface of the virion. Understanding of TMD dynamics is needed to better probe and target Env with small molecule and antibody therapies. However, little is known about TMD dynamics due to difficulties in describing native membrane properties. Here, we performed atomistic molecular dynamics simula- tions of a trimeric, prefusion TMD in a model, asymmetric viral membrane that mimics the native viral envelope. We found that water and chloride ions permeated the mem- brane and interacted with the highly conserved arginine bundle, (R696)3, at the center of the membrane and influenced TMD stability by creating a network of hydrogen bonds and electrostatic interactions.
    [Show full text]