Résumé
SARS-CoV-2, the virus responsible for the COVID-19 pandemic, encodes several accessory proteins, among which ORF6, a potent interferon inhibitor, is recognized as one of the most cytotoxic. Here, we investigated the structure, oligomeric state, and membrane interactions of ORF6 using NMR spectroscopy and molecular dynamics simulations. Using chemical-shift-ROSETTA, we show that ORF6 in proteoliposomes adopts a straight α-helical structure with an extended, rigid N-terminal part and flexible C-terminal residues. Cross-linking experiments indicate that ORF6 forms oligomers within lipid bilayers, and paramagnetic spin labeling suggests an antiparallel arrangement in its multimers. The amphipathic ORF6 helix establishes multiple contacts with the membrane surface with its N-terminal residues acting as membrane anchors. Our work demonstrates that ORF6 is an integral monotopic membrane protein and provides key insights into its conformation and the importance of the N-terminal region for the interaction with the membrane.