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TitleCryo-EM structure of SARS-CoV-2 ORF3a in lipid nanodiscs.
Journal, issue, pagesNat Struct Mol Biol, Vol. 28, Issue 7, Page 573-582, Year 2021
Publish dateJun 22, 2021
AuthorsDavid M Kern / Ben Sorum / Sonali S Mali / Christopher M Hoel / Savitha Sridharan / Jonathan P Remis / Daniel B Toso / Abhay Kotecha / Diana M Bautista / Stephen G Brohawn /
PubMed AbstractSARS-CoV-2 ORF3a is a putative viral ion channel implicated in autophagy inhibition, inflammasome activation and apoptosis. 3a protein and anti-3a antibodies are found in infected patient tissues and ...SARS-CoV-2 ORF3a is a putative viral ion channel implicated in autophagy inhibition, inflammasome activation and apoptosis. 3a protein and anti-3a antibodies are found in infected patient tissues and plasma. Deletion of 3a in SARS-CoV-1 reduces viral titer and morbidity in mice, suggesting it could be an effective target for vaccines or therapeutics. Here, we present structures of SARS-CoV-2 3a determined by cryo-EM to 2.1-Å resolution. 3a adopts a new fold with a polar cavity that opens to the cytosol and membrane through separate water- and lipid-filled openings. Hydrophilic grooves along outer helices could form ion-conduction paths. Using electrophysiology and fluorescent ion imaging of 3a-reconstituted liposomes, we observe Ca-permeable, nonselective cation channel activity, identify mutations that alter ion permeability and discover polycationic inhibitors of 3a activity. 3a-like proteins are found across coronavirus lineages that infect bats and humans, suggesting that 3a-targeted approaches could treat COVID-19 and other coronavirus diseases.
External linksNat Struct Mol Biol / PubMed:34158638 / PubMed Central
MethodsEM (single particle)
Resolution2.08 - 6.5 Å
Structure data

22136

6xdc

EMDB-22136, PDB-6xdc:
Cryo-EM structure of SARS-CoV-2 ORF3a
Method: EM (single particle) / Resolution: 2.9 Å

EMDB-22138:
Tetrameric SARS-CoV-2 ORF3a in a lipid nanodisc
Method: EM (single particle) / Resolution: 6.5 Å

EMDB-22139:
SARS-CoV-2 ORF3a with Emodin in a MSP1E3D1 lipid nanodisc
Method: EM (single particle) / Resolution: 3.7 Å

22898

7kjr

EMDB-22898, PDB-7kjr:
Cryo-EM structure of SARS-CoV-2 ORF3a
Method: EM (single particle) / Resolution: 2.08 Å

Chemicals

ChemComp-PEE:
1,2-dioleoyl-sn-glycero-3-phosphoethanolamine / DOPE, phospholipid*YM / Discrete optimized protein energy

ChemComp-HOH:
WATER / Water

Source
  • severe acute respiratory syndrome coronavirus 2
  • homo sapiens (human)
KeywordsTRANSPORT PROTEIN / SARS-CoV-2 / coronavirus / viroporin / ion channel / VIRAL PROTEIN / membrane protein

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