TY - JOUR

T1 - Activity of nsp14 Exonuclease from SARS-CoV-2 towards RNAs with Modified 3'-Termini

AU - Yuyukina, S K

AU - Barmatov, A E

AU - Bizyaev, S N

AU - Stetsenko, D A

AU - Sergeeva, O V

AU - Zatsepin, T S

AU - Zharkov, D O

N1 - Funding: This study was supported by the Russian Foundation for Basic Research (project no. 20-04-60433) and the Ministry of Science and Higher Education of the Russian Federation (project of Novosibirsk State University no. FSUS-2020-0035). © 2023. Pleiades Publishing, Ltd.

PY - 2023/4

Y1 - 2023/4

N2 - The COVID-19 pandemic has shown the urgent need for new treatments for coronavirus infections. Nucleoside analogs were successfully used to inhibit replication of some viruses through the incorporation into the growing DNA or RNA chain. However, the replicative machinery of coronaviruses contains nsp14, a non-structural protein with a 3'→5'-exonuclease activity that removes misincorporated and modified nucleotides from the 3' end of the growing RNA chain. Here, we studied the efficiency of hydrolysis of RNA containing various modifications in the 3'-terminal region by SARS-CoV-2 nsp14 exonuclease and its complex with the auxiliary protein nsp10. Single-stranded RNA was a preferable substrate compared to double-stranded RNA, which is consistent with the model of transfer of the substrate strand to the exonuclease active site, which was proposed on the basis of structural analysis. Modifications of the phosphodiester bond between the penultimate and last nucleotides had the greatest effect on nsp14 activity.

AB - The COVID-19 pandemic has shown the urgent need for new treatments for coronavirus infections. Nucleoside analogs were successfully used to inhibit replication of some viruses through the incorporation into the growing DNA or RNA chain. However, the replicative machinery of coronaviruses contains nsp14, a non-structural protein with a 3'→5'-exonuclease activity that removes misincorporated and modified nucleotides from the 3' end of the growing RNA chain. Here, we studied the efficiency of hydrolysis of RNA containing various modifications in the 3'-terminal region by SARS-CoV-2 nsp14 exonuclease and its complex with the auxiliary protein nsp10. Single-stranded RNA was a preferable substrate compared to double-stranded RNA, which is consistent with the model of transfer of the substrate strand to the exonuclease active site, which was proposed on the basis of structural analysis. Modifications of the phosphodiester bond between the penultimate and last nucleotides had the greatest effect on nsp14 activity.

KW - Humans

KW - SARS-CoV-2/genetics

KW - COVID-19

KW - Exonucleases

KW - Pandemics

KW - RNA, Viral/genetics

KW - Nucleotides

KW - Antiviral Agents/pharmacology

KW - Virus Replication/genetics

KW - RNA oligonucleotides

KW - antiviral drugs

KW - coronaviruses

KW - nsp14

KW - nsp10

KW - nucleoside inhibitors

KW - replication proofreading

KW - exonucleases

UR - https://www.scopus.com/record/display.uri?eid=2-s2.0-85162928320&origin=inward&txGid=e98eb6ba9e2a6745c1b0559cda11eca6

UR - https://www.mendeley.com/catalogue/c5023112-14db-3009-b341-c170f5a07492/

U2 - 10.1134/S1607672923700102

DO - 10.1134/S1607672923700102

M3 - Article

C2 - 37340295

VL - 509

SP - 65

EP - 69

JO - Doklady Biochemistry and Biophysics

JF - Doklady Biochemistry and Biophysics

SN - 1607-6729

IS - 1

ER -