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Principal Investigator: JAMES Jeiwen CHOU
Organization: BOSTON CHILDREN'S HOSPITAL
Fiscal Year: 2021
Award: $834,248
Funding agency: National Institute of Allergy and Infectious Diseases
Project Summary
The first critical step for enveloped viruses, such as HIV-1, to enter host cells is viral membrane fusion.
Viral fusion proteins are fascinating protein folding machineries capable of adopting completely different
conformations during the fusion process; they are also important vaccine and therapeutic targets. Previous
studies have revealed both pre- and post-fusion conformations of the soluble fragments of many viral fusion
proteins, but less is known for structures of their fusion peptide, transmembrane and membrane-proximal
regions in the context of lipid bilayers. There is strong evidence for functional roles of the membrane-
interacting regions in fusion, and yet mechanistic studies on how they exert their functions remain scarce. We
hypothesize that membrane-interacting regions of other fusion proteins related to HIV-1 envelope
protein (Env) adopt defined oligomeric structures that are critical for the stability, function and
antigenicity of the full-length proteins in membrane. In the studies that we completed during the previous
funding period, we have determined the structures of the TM, membrane proximal external region, and
cytoplasmic tail of HIV-1 Env in bicelles that mimic lipid bilayers using the latest NMR technology. We find that
these regions all form well-ordered trimeric clusters and are conformationally coupled, and that disrupting them
can reduce fusion and alter the antigenic structure of the entire Env. In this application, we propose to apply
our NMR/bicelle technology to investigate the membrane regions of SIV Env and the recently emerged SARS-
CoV-2 spike (S), and to use cryo-electron microscopy to determine structures of the full-length proteins
reconstituted in lipid nanodiscs. We will define roles in membrane fusion of critical structural elements of these
regions by deep mutagenesis and functional assays. We will purse the following specific aims: 1) we will
investigate the membrane-interacting components of SIV Env; 2) we will investigate the membrane-interacting
components in the postfusion arrangement; 3) we will determine structures of the full-length SIV Env and
SARS-CoV-2 S in the context of membrane; 4) we will elucidate roles of the membrane-interacting domains of
HIV/SIV Env and SARS-CoV-2 S in their stability, function and antigenicity.
Terms: <2019 novel corona virus><2019 novel coronavirus><2019-nCoV><2019-nCoV S protein><2019-nCoV spike glycoprotein><2019-nCoV spike protein><AIDS><AIDS Virus><Acquired Immune Deficiency><Acquired Immune Deficiency Syndrome><Acquired Immune Deficiency Syndrome Virus><Acquired Immuno-Deficiency Syndrome><Acquired Immunodeficiency Syndrome><Acquired Immunodeficiency Syndrome Virus><Acquired Immunologic Deficiency Syndrome><Adopted><Alanine><Animal Model><Animal Models and Related Studies><Architecture><Assay><Bioassay><Biogenesis><Biologic Assays><Biological Assay><COVID crisis><COVID epidemic><COVID pandemic><COVID-19 S protein><COVID-19 crisis><COVID-19 epidemic><COVID-19 global health crisis><COVID-19 global pandemic><COVID-19 health crisis><COVID-19 pandemic><COVID-19 public health crisis><COVID-19 spike glycoprotein><COVID-19 spike protein><COVID-19 virus><COVID19 S protein><COVID19 crisis><COVID19 epidemic><COVID19 global health crisis><COVID19 global pandemic><COVID19 health crisis><COVID19 pandemic><COVID19 public health crisis><COVID19 spike glycoprotein><COVID19 spike protein><COVID19 virus><Cell Body><Cell fusion><Cells><Chimera Protein><Chimeric Proteins><CoV-2><CoV2><Computer Models><Computerized Models><Coupled><Cryo-electron Microscopy><Cryoelectron Microscopy><Cytoplasmic Domain><Cytoplasmic Tail><Data Analyses><Data Analysis><Detergents><Disease><Disorder><Electron Cryomicroscopy><Elements><Engineering / Architecture><Envelope Protein><Environment><Funding><Fusion Protein><Genetics-Mutagenesis><Goals><HIV><HIV-1><HIV-I><HIV1><Human Immunodeficiency Virus Type 1><Human Immunodeficiency Viruses><Human immunodeficiency virus 1><Infection><Intervention><Intervention Strategies><LAV-HTLV-III><Length><Lipid Bilayers><Lipids><Lymphadenopathy-Associated Virus><M mulatta><M. mulatta><Macaca mulatta><Membrane><Membrane Fusion><Membrane Protein Gene><Membrane Proteins><Membrane-Associated Proteins><Molecular Configuration><Molecular Conformation><Molecular Stereochemistry><Mutagenesis><Mutagenesis Molecular Biology><Origin of Life><Peptides><Physiologic><Physiological><Play><Preparation><Process><Property><Proteins><Protocol><Protocols documentation><Research><Resolution><Rhesus Macaque><Rhesus Monkey><Role><SARS corona virus 2><SARS-CoV-2><SARS-CoV-2 S protein><SARS-CoV-2 epidemic><SARS-CoV-2 global health crisis><SARS-CoV-2 global pandemic><SARS-CoV-2 pandemic><SARS-CoV-2 spike glycoprotein><SARS-CoV-2 spike protein><SARS-CoV2><SARS-CoV2 S protein><SARS-CoV2 epidemic><SARS-CoV2 pandemic><SARS-CoV2 spike glycoprotein><SARS-CoV2 spike protein><SARS-associated corona virus 2><SARS-associated coronavirus 2><SARS-coronavirus-2><SARS-coronavirus-2 epidemic><SARS-coronavirus-2 pandemic><SARS-related corona virus 2><SARS-related coronavirus 2><SARSCoV2><SIV><Sampling><Scanning><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 CoV 2><Severe Acute Respiratory Syndrome CoV 2 epidemic><Severe Acute Respiratory Syndrome CoV 2 pandemic><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 corona virus 2><Severe acute respiratory syndrome coronavirus 2><Severe acute respiratory syndrome coronavirus 2 S protein><Severe acute respiratory syndrome coronavirus 2 epidemic><Severe acute respiratory syndrome coronavirus 2 pandemic><Severe acute respiratory syndrome coronavirus 2 spike glycoprotein><Severe acute respiratory syndrome coronavirus 2 spike protein><Severe acute respiratory syndrome related corona virus 2><Simian Immunodeficiency Viruses><Structure><Surface Proteins><TM Domain><Technology><Testing><Therapeutic><Transmembrane Domain><Transmembrane Region><Vaccines><Viral><Viral Diseases><Viral Fusion Proteins><Virus Assembly><Virus Diseases><Virus-HIV><Wuhan coronavirus><computational modeling><computational models><computer based models><computerized modeling><conformation><conformational state><corona virus disease 2019 epidemic><corona virus disease 2019 pandemic><coronavirus disease 2019 S protein><coronavirus disease 2019 crisis><coronavirus disease 2019 epidemic><coronavirus disease 2019 global health crisis><coronavirus disease 2019 global pandemic><coronavirus disease 2019 health crisis><coronavirus disease 2019 pandemic><coronavirus disease 2019 public health crisis><coronavirus disease 2019 spike glycoprotein><coronavirus disease 2019 spike protein><coronavirus disease 2019 virus><coronavirus disease crisis><coronavirus disease epidemic><coronavirus disease pandemic><cryo-EM><cryoEM><data interpretation><develop a vaccine><develop therapy><development of a vaccine><env Antigens><env Gene Products><env Polyproteins><env Protein><fascinate><fight against><hCoV19><insight><intervention development><interventional strategy><lipid bilayer membrane><membrane structure><model of animal><model organism><nCoV2><nano disk><nanodisc technology><nanodisk><novel><protein folding><protein structure><protein structures><proteins structure><reconstitute><reconstitution><severe acute respiratory syndrome coronavirus 2 global health crisis><severe acute respiratory syndrome coronavirus 2 global pandemic><social role><therapeutic agent development><therapeutic development><therapeutic target><therapy development><treatment development><vaccination study><vaccination trial><vaccine development><vaccine formulation><vaccine study><vaccine trial><viral assembly><viral infection><virus envelope><virus infection><virus-induced disease>