title: p53 down-regulates SARS coronavirus replication and is targeted by the SARS-unique domain and PLpro via E3 ubiquitin ligase RCHY1.
abstract: Highly pathogenic severe acute respiratory syndrome coronavirus (SARS-CoV) has developed strategies to inhibit host immune recognition. We identify cellular E3 ubiquitin ligase ring-finger and CHY zinc-finger domain-containing 1 (RCHY1) as an interacting partner of the viral SARS-unique domain (SUD) and papain-like protease (PL(pro)), and, as a consequence, the involvement of cellular p53 as antagonist of coronaviral replication. Residues 95-144 of RCHY1 and 389-652 of SUD (SUD-NM) subdomains are crucial for interaction. Association with SUD increases the stability of RCHY1 and augments RCHY1-mediated ubiquitination as well as degradation of p53. The calcium/calmodulin-dependent protein kinase II delta (CAMK2D), which normally influences RCHY1 stability by phosphorylation, also binds to SUD. In vivo phosphorylation shows that SUD does not regulate phosphorylation of RCHY1 via CAMK2D. Similarly to SUD, the PL(pro)s from SARS-CoV, MERS-CoV, and HCoV-NL63 physically interact with and stabilize RCHY1, and thus trigger degradation of endogenous p53. The SARS-CoV papain-like protease is encoded next to SUD within nonstructural protein 3. A SUD-PL(pro) fusion interacts with RCHY1 more intensively and causes stronger p53 degradation than SARS-CoV PL(pro) alone. We show that p53 inhibits replication of infectious SARS-CoV as well as of replicons and human coronavirus NL63. Hence, human coronaviruses antagonize the viral inhibitor p53 via stabilizing RCHY1 and promoting RCHY1-mediated p53 degradation. SUD functions as an enhancer to strengthen interaction between RCHY1 and nonstructural protein 3, leading to a further increase in in p53 degradation. The significance of these findings is that down-regulation of p53 as a major player in antiviral innate immunity provides a long-sought explanation for delayed activities of respective genes.
authors: Institute of Biochemistry, Center for Structural and Cell Biology in Medicine, University of Lübeck and German Center for Infection Research, partner site Luebeck, 23538 Luebeck, Germany; Max-von-Pettenkofer Institute, Ludwig-Maximilians-University Munich and German Center for Infection Research (DZIF), partner site Munich, 80336 Munich, Germany;
authors: Yue
authors: Y
authors: Ma-Lauer
authors: Max-von-Pettenkofer Institute, Ludwig-Maximilians-University Munich and German Center for Infection Research (DZIF), partner site Munich, 80336 Munich, Germany;
authors: Javier
authors: J
authors: Carbajo-Lozoya
authors: Department of Proteomics and Signal Transduction, Max-Planck Institute of Biochemistry, 82152 Martinsried, Germany;
authors: Marco Y
authors: 0000-0002-9490-2261
authors: MY
authors: Hein
authors: Institute of Virology, University of Bonn Medical Centre and German Center for Infection Research, partner site Bonn, 53127 Bonn, Germany;
authors: Marcel A
authors: MA
authors: Müller
authors: Department of Biology and Center for Integrated Protein Science, Ludwig-Maximilians-University Munich, 82152 Planegg-Martinsried, Germany;
authors: Wen
authors: W
authors: Deng
authors: Institute of Biochemistry, Center for Structural and Cell Biology in Medicine, University of Lübeck and German Center for Infection Research, partner site Luebeck, 23538 Luebeck, Germany;
authors: Jian
authors: J
authors: Lei
authors: Institute of Virology, University of Bonn Medical Centre and German Center for Infection Research, partner site Bonn, 53127 Bonn, Germany;
authors: Benjamin
authors: B
authors: Meyer
authors: Institute of Biochemistry, Center for Structural and Cell Biology in Medicine, University of Lübeck and German Center for Infection Research, partner site Luebeck, 23538 Luebeck, Germany;
authors: Yuri
authors: Y
authors: Kusov
authors: Max-von-Pettenkofer Institute, Ludwig-Maximilians-University Munich and German Center for Infection Research (DZIF), partner site Munich, 80336 Munich, Germany;
authors: Brigitte
authors: B
authors: von Brunn
authors: Max-von-Pettenkofer Institute, Ludwig-Maximilians-University Munich and German Center for Infection Research (DZIF), partner site Munich, 80336 Munich, Germany;
authors: Dev Raj
authors: DR
authors: Bairad
authors: Experimental and Molecular Pathology, Institute of Pathology, Ludwig-Maximilians-University Munich, 80337 Munich, Germany.
authors: Sabine
authors: S
authors: Hünten
authors: Institute of Virology, University of Bonn Medical Centre and German Center for Infection Research, partner site Bonn, 53127 Bonn, Germany;
authors: Christian
authors: C
authors: Drosten
authors: Experimental and Molecular Pathology, Institute of Pathology, Ludwig-Maximilians-University Munich, 80337 Munich, Germany.
authors: Heiko
authors: H
authors: Hermeking
authors: Department of Biology and Center for Integrated Protein Science, Ludwig-Maximilians-University Munich, 82152 Planegg-Martinsried, Germany;
authors: Heinrich
authors: H
authors: Leonhardt
authors: Department of Proteomics and Signal Transduction, Max-Planck Institute of Biochemistry, 82152 Martinsried, Germany;
authors: Matthias
authors: M
authors: Mann
authors: Institute of Biochemistry, Center for Structural and Cell Biology in Medicine, University of Lübeck and German Center for Infection Research, partner site Luebeck, 23538 Luebeck, Germany;
authors: Rolf
authors: R
authors: Hilgenfeld
authors: Max-von-Pettenkofer Institute, Ludwig-Maximilians-University Munich and German Center for Infection Research (DZIF), partner site Munich, 80336 Munich, Germany; vonbrunn@mvp.uni-muenchen.de.
authors: Albrecht
authors: A
authors: von Brunn
mesh_terms: D001665:Binding Sites / Q000235:genetics; D054732:Calcium-Calmodulin-Dependent Protein Kinase Type 2 / Q000235:genetics / Q000378:metabolism; D000086782:Coronavirus 3C Proteases; D003546:Cysteine Endopeptidases / Q000235:genetics / Q000378:metabolism*; D015536:Down-Regulation; D054884:Host-Pathogen Interactions; D006801:Humans; D011485:Protein Binding; D045473:Severe acute respiratory syndrome-related coronavirus / Q000235:genetics / Q000378:metabolism* / Q000502:physiology; D045169:Severe Acute Respiratory Syndrome / Q000235:genetics / Q000378:metabolism* / Q000821:virology; D016159:Tumor Suppressor Protein p53 / Q000378:metabolism*; D044767:Ubiquitin-Protein Ligases / Q000235:genetics / Q000378:metabolism*; D054875:Ubiquitination; D017361:Viral Nonstructural Proteins / Q000235:genetics / Q000378:metabolism; D014764:Viral Proteins / Q000235:genetics / Q000378:metabolism*; D014779:Virus Replication / Q000235:genetics
chemical_list: C000595202:Nsp3 protein, Middle East respiratory syndrome coronavirus; D016159:Tumor Suppressor Protein p53; D017361:Viral Nonstructural Proteins; D014764:Viral Proteins; C485369:RCHY1 protein, human; D044767:Ubiquitin-Protein Ligases; C517660:CAMK2D protein, human; D054732:Calcium-Calmodulin-Dependent Protein Kinase Type 2; D003546:Cysteine Endopeptidases; D000086782:Coronavirus 3C Proteases
keywords: E3 ubiquitin ligase RCHY1; SARS-CoV SUD; coronavirus replication; p53 antiviral activity; papain-like protease