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Principal Investigator: Ricardo Rajsbaum
Organization: RUTGERS BIOMEDICAL AND HEALTH SCIENCES
Fiscal Year: 2024
Award: $471,011
Funding agency: National Institute of Allergy and Infectious Diseases
The Severe Acute Respiratory Syndrome Coronavirus 2 (CoV-2) belongs to a family of pathogenic enveloped RNA viruses of the family Coronaviridae. The ongoing pandemic has caused a public health emergency worldwide, accompanied by dire health and economic consequences. There is evidence suggesting that CoV-2 may have relatively higher infection rates compared to previous epidemic strains of SARS and higher affinity to the receptor ACE2 than SARS-1. In addition, new CoV-2 variants have appeared recently with mutations that correlate with higher infection rates and the ability to escape specific immunity, causing major concerns. A major gap in knowledge remains as to how CoV-2 may have acquired the ability to spread more efficiently, and how new mutations may affect virus infectivity. The overarching goal of this proposal is to better understand the molecular mechanisms that regulate CoV-2 cell entry and replication, and how the appearance of new variants could lead to immune escape. We will focus on the role of the host Ubiquitin (Ub) system in promoting CoV-2 infection. This information could help predict appearance of more transmissible variants of coronaviruses, and to develop antiviral approaches by targeting specific steps of the ubiquitination process. Our data recently published in Nature, show that the envelope protein of flaviviruses is K63-linked polyubiquitinated, which enhances virus attachment to host cell receptors. Therefore, we asked whether a similar mechanism applies to SARS-CoV-2. Our preliminary data indicate that CoV-2 structural proteins are ubiquitinated on multiple lysine residue, some of which are not conserved in the original epidemic CoV strain. In addition, new variants of CoV-2 have appeared with mutations on these ubiquitination sites. Our data also suggest that ubiquitination of Spike (S) protein may play a role in stabilizing the CoV-2 S-ACE2 interaction, potentially leading to enhanced entry and pathogenesis. It is currently unknown whether any member of the Coronaviridae family, including SARS-CoV-2, utilize ubiquitination of viral structural proteins as a mechanism of virus attachment and entry. We have also identified E3-Ub ligases of the Tripartite Motif (TRIM) family of proteins, which ubiquitinates viral structural proteins. Our general hypothesis is that the variants of CoV-2 that have gained specific lysine residues provide new Ub acceptor sites on structural proteins, which can enhance virus replication and immune escape. By using in vitro biochemical approaches, novel recombinant mutant viruses, and in vivo models, we will assess how ubiquitination of structural CoV-2 proteins contribute to CoV-2 infectivity. In Aim 1 we will determine the mechanistic role of ubiquitination of the CoV-2 S protein in virus replication and antibody escape, and in Aim 2 we will determine the mechanistic role of ubiquitination of the CoV-2 Membrane protein in virus replication and IFN antagonism. The outcome of these studies may help explain how new more infectious viruses may appear by gaining ubiquitination sites and will provide the basis for the development of an antiviral approach that could be applied to a broad range of enveloped viruses.
Terms: <2019 novel corona virus><2019 novel coronavirus><2019-nCoV><2019-nCoV S protein><2019-nCoV spike glycoprotein><2019-nCoV spike protein><2019-nCoV variant><2019-nCoV variant forms><2019-nCoV variant strains><ACE2><APF-1><ATP-Dependent Proteolysis Factor 1><Affect><Affinity><Antibodies><Appearance><Assay><BSL-3 facility><BSL3 facility><Binding><Bio-Informatics><Bioassay><Biochemical><Bioinformatics><Biological Assay><Blocking Antibodies><COVID-19><COVID-19 S protein><COVID-19 infection><COVID-19 spike><COVID-19 spike glycoprotein><COVID-19 spike protein><COVID-19 variant><COVID-19 variant forms><COVID-19 variant strains><COVID-19 virus><COVID-19 virus infection><COVID19 infection><COVID19 virus><CV-19><Cell Body><Cell Line><CellLine><Cells><Cessation of life><Clinical Trials><Co-Immunoprecipitations><CoV emergence><CoV-2><CoV2><Coronaviridae><Coronavirus><Coronavirus Infectious Disease 2019><Data><Death><Disease><Disorder><Economics><Envelope Protein><Epidemic><Family><Flaviviridae><Flavivirus><Future><Genetic Alteration><Genetic Change><Genetic defect><Genetics-Mutagenesis><Goals><Group B Arbovirus><HMG-20><Health><High Mobility Protein 20><Host Factor><Host Factor Protein><Human><IFN><IFN antagonist><Immune><Immune Evasion><Immunes><Immunity><Impairment><In Vitro><Integration Host Factors><Interferons><Knowledge><L-Lysine><Ligase><Ligase Gene><Link><Lung Diseases><Lysine><M protein><Mass Photometry/Spectrum Analysis><Mass Spectrometry><Mass Spectroscopy><Mass Spectrum><Mass Spectrum Analyses><Mass Spectrum Analysis><Mediating><Membrane Protein Gene><Membrane Proteins><Membrane-Associated Proteins><Mice><Mice Mammals><Modern Man><Molecular><Molecular Interaction><Murine><Mus><Mutagenesis><Mutagenesis Molecular Biology><Mutation><Names><Nature><Orthoflavivirus><Outcome><Outcome Study><Pathogenesis><Pathogenicity><Play><Process><Protein Family><Proteins><Publishing><Pulmonary Diseases><Pulmonary Disorder><RBCC/TRIM Motif><RNA Viruses><Receptor Cell><Receptor Protein><Recombinants><Reporting><Role><SARS><SARS Virus><SARS corona virus><SARS corona virus 2><SARS coronavirus><SARS coronavirus disease><SARS-Associated Coronavirus><SARS-CO-V2><SARS-COVID-2><SARS-CoV><SARS-CoV disease><SARS-CoV-1><SARS-CoV-2><SARS-CoV-2 S><SARS-CoV-2 S protein><SARS-CoV-2 infection><SARS-CoV-2 spike><SARS-CoV-2 spike glycoprotein><SARS-CoV-2 spike protein><SARS-CoV-2 variant><SARS-CoV-2 variant forms><SARS-CoV-2 variant strains><SARS-CoV2><SARS-CoV2 infection><SARS-Related Coronavirus><SARS-associated corona virus 2><SARS-associated coronavirus 2><SARS-coronavirus-2><SARS-related corona virus 2><SARS-related coronavirus 2><SARSCoV2><Severe Acute Respiratory Coronavirus><Severe Acute Respiratory Coronavirus 2><Severe Acute Respiratory Distress Syndrome CoV 2><Severe Acute Respiratory Distress Syndrome Corona Virus 2><Severe Acute Respiratory Distress Syndrome Coronavirus 2><Severe Acute Respiratory Syndrome><Severe Acute Respiratory Syndrome CoV 2><Severe Acute Respiratory Syndrome CoV disease><Severe Acute Respiratory Syndrome Virus><Severe Acute Respiratory Syndrome corona virus><Severe Acute Respiratory Syndrome coronavirus><Severe Acute Respiratory Syndrome coronavirus disease><Severe Acute Respiratory Syndrome-associated coronavirus 2><Severe Acute Respiratory Syndrome-related coronavirus 2><Severe acute respiratory syndrome associated corona virus 2><Severe acute respiratory syndrome coronavirus 2><Severe acute respiratory syndrome coronavirus 2 S protein><Severe acute respiratory syndrome coronavirus 2 infection><Severe acute respiratory syndrome coronavirus 2 spike glycoprotein><Severe acute respiratory syndrome coronavirus 2 spike protein><Severe acute respiratory syndrome related corona virus 2><Site><Strains Cell Lines><Structural Protein><Surface Proteins><Synthetases><System><TRIM Motif><Testing><Therapeutic Intervention><Time><Transmission><Tripartite Motif><Ubiquitilation><Ubiquitin><Ubiquitination><Ubiquitinoylation><Vaccines><Variant><Variation><Viral><Viral M Proteins><Viral Matrix Proteins><Viral Membrane Proteins><Viral Packaging><Viral Receptor><Viral Structural Proteins><Virion><Virus><Virus Packagings><Virus Particle><Virus Receptors><Virus Replication><Wuhan coronavirus><angiotensin converting enzyme 2><angiotensin converting enzyme II><antagonism><antagonist><anti-viral development><anti-viral drug development><anti-viral therapeutic development><anti-viral therapy development><antiviral development><antiviral drug development><antiviral therapeutic development><antiviral therapy development><biosafety level 3 facility><corona virus><corona virus emergence><coronavirus disease 2019><coronavirus disease 2019 S protein><coronavirus disease 2019 infection><coronavirus disease 2019 spike glycoprotein><coronavirus disease 2019 spike protein><coronavirus disease 2019 variant><coronavirus disease 2019 variant forms><coronavirus disease 2019 variant strains><coronavirus disease 2019 virus><coronavirus disease-19><coronavirus disease-19 virus><coronavirus emergence><coronavirus infectious disease-19><cultured cell line><developing anti-viral agent><developing anti-viral drug><developing anti-viral therapeutic><developing anti-viral therapy><developing antiviral agent><developing antiviral drug><developing antiviral therapeutic><developing antiviral therapy><disease of the lung><disorder of the lung><economic><emergent CoV><emergent corona virus><emergent coronavirus><emerging CoV><emerging corona virus><emerging coronavirus><env Antigens><env Gene Products><env Polyproteins><env Protein><genome mutation><hCoV19><immune evasive><in vivo><in vivo Model><infected with COVID-19><infected with COVID19><infected with SARS-CoV-2><infected with SARS-CoV2><infected with coronavirus disease 2019><infected with severe acute respiratory syndrome coronavirus 2><infection rate><interferon antagonist><intervention therapy><lung disorder><member><multiple myeloma M Protein><mutant><nCoV><nCoV2><name><named><naming><neutralizing antibody><new CoV><new corona virus><new coronavirus><novel><novel CoV><novel corona virus><novel coronavirus><pandemic><pandemic disease><past epidemic><pathogenic virus><pharmacologic><previous epidemic><prior epidemic><public health emergency><rate of infection><receptor><severe acute respiratory syndrome coronavirus 2 variant><severe acute respiratory syndrome coronavirus 2 variant forms><severe acute respiratory syndrome coronavirus 2 variant strains><severe acute respiratory syndrome-CoV><social role><spike proteins on SARS-CoV-2><targeted drug therapy><targeted drug treatments><targeted therapeutic><targeted therapeutic agents><targeted therapy><targeted treatment><transmission process><ubiquination><ubiquitin conjugation><viral multiplication><viral pathogen><viral replication><virus multiplication><virus pathogen>