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Fidelity of Ribonucleotide Incorporation by the SARS-CoV-2 Replication Complex.

Xingyu Yin1, Horia Popa1, Anthony Stapon1

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|January 23, 2023
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Summary
This summary is machine-generated.

SARS-CoV-2 RNA replication fidelity is surprisingly low without proofreading. Mutations in key viral proteins nsp12 and nsp14 may further reduce accuracy, impacting viral evolution and vaccine development.

Keywords:
RNA-dependent RNA polymeraseSARS-CoV-2exoribonucleasemutationviral replication fidelity

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Area of Science:

  • Virology
  • Molecular Biology
  • Genetics

Background:

  • The SARS-CoV-2 pandemic is driven by viral genomic variation.
  • Understanding SARS-CoV-2 replication fidelity is crucial due to mutations.
  • RNA-dependent RNA polymerase (RdRp) is central to viral replication.

Purpose of the Study:

  • To determine the fidelity of SARS-CoV-2 RdRp (nsp12) with its co-factors (nsp7/nsp8).
  • To investigate the impact of mutations in nsp12 and nsp14 on replication fidelity.

Main Methods:

  • Steady-state kinetic analysis of ribonucleotide incorporation by the nsp12/7/8 complex.
  • Mutational analysis of reported SARS-CoV-2 genomes.

Main Results:

  • The nsp12/7/8 complex exhibits low base substitution fidelity (10^-1-10^-3) without proofreading (nsp14).
  • This fidelity is significantly lower than other coronaviruses (10^-6-10^-7).
  • Mutations in nsp12 and nsp14 were identified that likely impair proofreading and reduce replication fidelity.

Conclusions:

  • Proofreading by nsp14 is essential for high-fidelity SARS-CoV-2 RNA replication.
  • Mutations affecting nsp12 and nsp14 function can lower viral replication fidelity.
  • These findings inform the development of antivirals and vaccines targeting SARS-CoV-2.