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Structural basis for backtracking by the SARS-CoV-2 replication-transcription complex.

Brandon Malone1,2, James Chen1,2, Qi Wang3

  • 1Laboratory of Molecular Biophysics, The Rockefeller University, New York, NY, 10065 USA.

Biorxiv : the Preprint Server for Biology
|March 24, 2021
PubMed
Summary
This summary is machine-generated.

Backtracking, the reverse motion of RNA-dependent RNA polymerase (RdRp), is crucial for SARS-CoV-2 transcription and replication. This viral backtracking mechanism may enhance proofreading, impacting antiviral resistance.

Keywords:
Biological SciencesBiophysics and Computational BiologyRNA-dependent RNA polymerasebacktrackingcoronaviruscryo-electron microscopymolecular dynamics

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Area of Science:

  • Virology
  • Molecular Biology
  • Structural Biology

Background:

  • Backtracking is a known regulatory mechanism in cellular transcription but its role in viral transcription is unclear.
  • Understanding viral backtracking is essential for deciphering viral replication strategies and developing antivirals.

Approach:

  • Utilized cryo-electron microscopy, RNA-protein crosslinking, and molecular dynamics simulations.
  • Investigated the interaction between SARS-CoV-2 RNA-dependent RNA polymerase (RdRp) and the nsp13 helicase.
  • Characterized the structural and mechanistic aspects of SARS-CoV-2 RdRp backtracking.

Key Points:

  • SARS-CoV-2 RdRp backtracking extrudes product-RNA through the NTP-entry tunnel.
  • Mismatched nucleotides at the 3'-end of product-RNA initiate backtracking by entering the NTP-entry tunnel.
  • The nsp13 helicase significantly stimulates SARS-CoV-2 RdRp backtracking.

Conclusions:

  • Backtracking is a functional mechanism in SARS-CoV-2 transcription and replication.
  • This process may contribute to viral proofreading, influencing resistance to antiviral therapies.
  • The findings provide insights into viral polymerase dynamics and potential therapeutic targets.