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Ensemble cryo-electron microscopy reveals conformational states of the nsp13 helicase in the SARS-CoV-2 helicase replication-transcription complex
James Chen; Qi Wang; Brandon Malone; Eliza Llewellyn; Yakov Pechersky; Kashyap Maruthi; Edward T Eng; Jason K Perry; Elizabeth A. Campbell; David E. Shaw; Seth A Darst.
Afiliación
  • James Chen; The Rockefeller University
  • Qi Wang; D. E. Shaw Research
  • Brandon Malone; The Rockefeller University
  • Eliza Llewellyn; The Rockefeller University
  • Yakov Pechersky; D. E. Shaw Research
  • Kashyap Maruthi; The National Resource for Automated Molecular Microscopy, Simons Electron Microscopy Center, New York Structural Biology Center
  • Edward T Eng; New York Structural Biology Center
  • Jason K Perry; Gilead Sciences
  • Elizabeth A. Campbell; The Rockefeller University
  • David E. Shaw; D. E. Shaw Research
  • Seth A Darst; The Rockefeller University
Preprint en En | PREPRINT-BIORXIV | ID: ppbiorxiv-468168
ABSTRACT
The SARS-CoV-2 nonstructural proteins coordinate genome replication and gene expression. Structural analyses revealed the basis for coupling of the essential nsp13 helicase with the RNA dependent RNA polymerase (RdRp) where the holo-RdRp and RNA substrate (the replication-transcription complex, or RTC) associated with two copies of nsp13 (nsp132-RTC). One copy of nsp13 interacts with the template RNA in an opposing polarity to the RdRp and is envisaged to drive the RdRp backwards on the RNA template (backtracking), prompting questions as to how the RdRp can efficiently synthesize RNA in the presence of nsp13. Here, we use cryo-electron microscopy and molecular dynamics simulations to analyze the nsp132-RTC, revealing four distinct conformational states of the helicases. The results suggest a mechanism for the nsp132-RTC to turn backtracking on and off, using an allosteric mechanism to switch between RNA synthesis or backtracking in response to stimuli at the RdRp active site.
Licencia
cc_by_nc_nd
Texto completo: 1 Colección: 09-preprints Base de datos: PREPRINT-BIORXIV Idioma: En Año: 2021 Tipo del documento: Preprint
Texto completo: 1 Colección: 09-preprints Base de datos: PREPRINT-BIORXIV Idioma: En Año: 2021 Tipo del documento: Preprint