Biochemical characterization of a recombinant SARS coronavirus nsp12 RNA-dependent RNA polymerase capable of copying viral RNA templates

Dae Gyun Ahn, Jin Kyu Choi, Deborah R. Taylor, Jong Won Oh

Research output: Contribution to journalArticle

15 Citations (Scopus)

Abstract

The severe acute respiratory syndrome coronavirus (SARS-CoV) RNA genome is replicated by a virus-encoded RNA replicase, the key component of which is the nonstructural protein 12 (nsp12). In this report, we describe the biochemical properties of a full-length recombinant SARS-CoV nsp12 RNA-dependent RNA polymerase (RdRp) capable of copying viral RNA templates. The purified SARS-CoV nsp12 showed both primer-dependent and primer-independent RNA synthesis activities using homopolymeric RNA templates. The RdRp activity was strictly dependent on Mn2+. The nsp12 preferentially copied homopolymeric pyrimidine RNA templates in the absence of an added oligonucleotide primer. It was also able to initiate de novo RNA synthesis from the 3'-ends of both the plus- and minus-strand genome of SARS-CoV, using the 3'-terminal 36- and 37-nt RNA, respectively. The in vitro RdRp assay system established with a full-length nsp12 will be useful for understanding the mechanisms of coronavirus replication and for the development of anti-SARS-CoV agents.

Original languageEnglish
Pages (from-to)2095-2104
Number of pages10
JournalArchives of Virology
Volume157
Issue number11
DOIs
Publication statusPublished - 2012 Nov 1

Fingerprint

SARS Virus
RNA Replicase
Severe Acute Respiratory Syndrome
Viral RNA
Coronavirus
RNA
Genome
Proteins
DNA Primers
coronavirus nonstructural protein
Viruses

All Science Journal Classification (ASJC) codes

  • Virology

Cite this

@article{3afe7a68b35c49cba656f06584d266dd,
title = "Biochemical characterization of a recombinant SARS coronavirus nsp12 RNA-dependent RNA polymerase capable of copying viral RNA templates",
abstract = "The severe acute respiratory syndrome coronavirus (SARS-CoV) RNA genome is replicated by a virus-encoded RNA replicase, the key component of which is the nonstructural protein 12 (nsp12). In this report, we describe the biochemical properties of a full-length recombinant SARS-CoV nsp12 RNA-dependent RNA polymerase (RdRp) capable of copying viral RNA templates. The purified SARS-CoV nsp12 showed both primer-dependent and primer-independent RNA synthesis activities using homopolymeric RNA templates. The RdRp activity was strictly dependent on Mn2+. The nsp12 preferentially copied homopolymeric pyrimidine RNA templates in the absence of an added oligonucleotide primer. It was also able to initiate de novo RNA synthesis from the 3'-ends of both the plus- and minus-strand genome of SARS-CoV, using the 3'-terminal 36- and 37-nt RNA, respectively. The in vitro RdRp assay system established with a full-length nsp12 will be useful for understanding the mechanisms of coronavirus replication and for the development of anti-SARS-CoV agents.",
author = "Ahn, {Dae Gyun} and Choi, {Jin Kyu} and Taylor, {Deborah R.} and Oh, {Jong Won}",
year = "2012",
month = "11",
day = "1",
doi = "10.1007/s00705-012-1404-x",
language = "English",
volume = "157",
pages = "2095--2104",
journal = "Archives of Virology",
issn = "0304-8608",
publisher = "Springer Wien",
number = "11",

}

Biochemical characterization of a recombinant SARS coronavirus nsp12 RNA-dependent RNA polymerase capable of copying viral RNA templates. / Ahn, Dae Gyun; Choi, Jin Kyu; Taylor, Deborah R.; Oh, Jong Won.

In: Archives of Virology, Vol. 157, No. 11, 01.11.2012, p. 2095-2104.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Biochemical characterization of a recombinant SARS coronavirus nsp12 RNA-dependent RNA polymerase capable of copying viral RNA templates

AU - Ahn, Dae Gyun

AU - Choi, Jin Kyu

AU - Taylor, Deborah R.

AU - Oh, Jong Won

PY - 2012/11/1

Y1 - 2012/11/1

N2 - The severe acute respiratory syndrome coronavirus (SARS-CoV) RNA genome is replicated by a virus-encoded RNA replicase, the key component of which is the nonstructural protein 12 (nsp12). In this report, we describe the biochemical properties of a full-length recombinant SARS-CoV nsp12 RNA-dependent RNA polymerase (RdRp) capable of copying viral RNA templates. The purified SARS-CoV nsp12 showed both primer-dependent and primer-independent RNA synthesis activities using homopolymeric RNA templates. The RdRp activity was strictly dependent on Mn2+. The nsp12 preferentially copied homopolymeric pyrimidine RNA templates in the absence of an added oligonucleotide primer. It was also able to initiate de novo RNA synthesis from the 3'-ends of both the plus- and minus-strand genome of SARS-CoV, using the 3'-terminal 36- and 37-nt RNA, respectively. The in vitro RdRp assay system established with a full-length nsp12 will be useful for understanding the mechanisms of coronavirus replication and for the development of anti-SARS-CoV agents.

AB - The severe acute respiratory syndrome coronavirus (SARS-CoV) RNA genome is replicated by a virus-encoded RNA replicase, the key component of which is the nonstructural protein 12 (nsp12). In this report, we describe the biochemical properties of a full-length recombinant SARS-CoV nsp12 RNA-dependent RNA polymerase (RdRp) capable of copying viral RNA templates. The purified SARS-CoV nsp12 showed both primer-dependent and primer-independent RNA synthesis activities using homopolymeric RNA templates. The RdRp activity was strictly dependent on Mn2+. The nsp12 preferentially copied homopolymeric pyrimidine RNA templates in the absence of an added oligonucleotide primer. It was also able to initiate de novo RNA synthesis from the 3'-ends of both the plus- and minus-strand genome of SARS-CoV, using the 3'-terminal 36- and 37-nt RNA, respectively. The in vitro RdRp assay system established with a full-length nsp12 will be useful for understanding the mechanisms of coronavirus replication and for the development of anti-SARS-CoV agents.

UR - http://www.scopus.com/inward/record.url?scp=84868492015&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84868492015&partnerID=8YFLogxK

U2 - 10.1007/s00705-012-1404-x

DO - 10.1007/s00705-012-1404-x

M3 - Article

C2 - 22791111

AN - SCOPUS:84868492015

VL - 157

SP - 2095

EP - 2104

JO - Archives of Virology

JF - Archives of Virology

SN - 0304-8608

IS - 11

ER -