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Article: The severe acute respiratory syndrome (SARS) coronavirus NTPasefhelicase belongs to a distinct class of 5′ to 3′ viral helicases

TitleThe severe acute respiratory syndrome (SARS) coronavirus NTPasefhelicase belongs to a distinct class of 5′ to 3′ viral helicases
Authors
Issue Date2003
PublisherAmerican Society for Biochemistry and Molecular Biology, Inc. The Journal's web site is located at http://www.jbc.org/
Citation
Journal Of Biological Chemistry, 2003, v. 278 n. 41, p. 39578-39582 How to Cite?
AbstractThe putative NTPase/helicase protein from severe acute respiratory syndrome coronavirus (SARS-CoV) is postulated to play a number of crucial roles in the viral life cycle, making it an attractive target for anti-SARS therapy. We have cloned, expressed, and purified this protein as an N-terminal hexahistidine fusion in Escherichia coli and have characterized its helicase and NTPase activities. The enzyme unwinds double-stranded DNA, dependent on the presence of a 5′ single-stranded overhang, indicating a 5′ to 3′ polarity of activity, a distinct characteristic of coronaviridae helicases. We provide the first quantitative analysis of the polynucleic acid binding and NTPase activities of a Nidovirus helicase, using a high throughput phosphate release assay that will be readily adaptable to the future testing of helicase inhibitors. All eight common NTPs and dNTPs were hydrolyzed by the SARS helicase in a magnesium-dependent reaction, stimulated by the presence of either single-stranded DNA or RNA. The enzyme exhibited a preference for ATP, dATP, and dCTP over the other NTP/dNTP substrates. Homopolynucleotides significantly stimulated the ATPase activity (15-25-fold) with the notable exception of poly(G) and poly(dG), which were non-stimulatory. We found a large variation in the apparent strength of binding of different homopolynucleotides, with dT 24 binding over 10 times more strongly than dA 24 as observed by the apparent K m.
Persistent Identifierhttp://hdl.handle.net/10722/147488
ISSN
2015 Impact Factor: 4.258
2015 SCImago Journal Rankings: 3.151
ISI Accession Number ID
References

 

DC FieldValueLanguage
dc.contributor.authorTanner, JAen_HK
dc.contributor.authorWatt, RMen_HK
dc.contributor.authorChai, YBen_HK
dc.contributor.authorLu, LYen_HK
dc.contributor.authorLin, MCen_HK
dc.contributor.authorPeiris, JSMen_HK
dc.contributor.authorPoon, LLMen_HK
dc.contributor.authorKung, HFen_HK
dc.contributor.authorHuang, JDen_HK
dc.date.accessioned2012-05-29T06:04:04Z-
dc.date.available2012-05-29T06:04:04Z-
dc.date.issued2003en_HK
dc.identifier.citationJournal Of Biological Chemistry, 2003, v. 278 n. 41, p. 39578-39582en_HK
dc.identifier.issn0021-9258en_HK
dc.identifier.urihttp://hdl.handle.net/10722/147488-
dc.description.abstractThe putative NTPase/helicase protein from severe acute respiratory syndrome coronavirus (SARS-CoV) is postulated to play a number of crucial roles in the viral life cycle, making it an attractive target for anti-SARS therapy. We have cloned, expressed, and purified this protein as an N-terminal hexahistidine fusion in Escherichia coli and have characterized its helicase and NTPase activities. The enzyme unwinds double-stranded DNA, dependent on the presence of a 5′ single-stranded overhang, indicating a 5′ to 3′ polarity of activity, a distinct characteristic of coronaviridae helicases. We provide the first quantitative analysis of the polynucleic acid binding and NTPase activities of a Nidovirus helicase, using a high throughput phosphate release assay that will be readily adaptable to the future testing of helicase inhibitors. All eight common NTPs and dNTPs were hydrolyzed by the SARS helicase in a magnesium-dependent reaction, stimulated by the presence of either single-stranded DNA or RNA. The enzyme exhibited a preference for ATP, dATP, and dCTP over the other NTP/dNTP substrates. Homopolynucleotides significantly stimulated the ATPase activity (15-25-fold) with the notable exception of poly(G) and poly(dG), which were non-stimulatory. We found a large variation in the apparent strength of binding of different homopolynucleotides, with dT 24 binding over 10 times more strongly than dA 24 as observed by the apparent K m.en_HK
dc.languageengen_US
dc.publisherAmerican Society for Biochemistry and Molecular Biology, Inc. The Journal's web site is located at http://www.jbc.org/en_HK
dc.relation.ispartofJournal of Biological Chemistryen_HK
dc.subject.meshAnimalsen_US
dc.subject.meshBase Sequenceen_US
dc.subject.meshCercopithecus Aethiopsen_US
dc.subject.meshDna Helicases - Classification - Genetics - Metabolismen_US
dc.subject.meshDna, Viral - Genetics - Metabolismen_US
dc.subject.meshKineticsen_US
dc.subject.meshMolecular Sequence Dataen_US
dc.subject.meshNucleoside-Triphosphatase - Classification - Genetics - Metabolismen_US
dc.subject.meshRna Helicases - Classification - Genetics - Metabolismen_US
dc.subject.meshSars Virus - Enzymology - Geneticsen_US
dc.subject.meshSubstrate Specificityen_US
dc.subject.meshVero Cellsen_US
dc.titleThe severe acute respiratory syndrome (SARS) coronavirus NTPasefhelicase belongs to a distinct class of 5′ to 3′ viral helicasesen_HK
dc.typeArticleen_HK
dc.identifier.emailTanner, JA: jatanner@hku.hken_HK
dc.identifier.emailWatt, RM: rmwatt@hku.hken_HK
dc.identifier.emailLin, MC: mcllin@hkucc.hku.hken_HK
dc.identifier.emailPeiris, JSM: malik@hkucc.hku.hken_HK
dc.identifier.emailPoon, LLM: llmpoon@hkucc.hku.hken_HK
dc.identifier.emailHuang, JD: jdhuang@hku.hken_HK
dc.identifier.authorityTanner, JA=rp00495en_HK
dc.identifier.authorityWatt, RM=rp00043en_HK
dc.identifier.authorityLin, MC=rp00746en_HK
dc.identifier.authorityPeiris, JSM=rp00410en_HK
dc.identifier.authorityPoon, LLM=rp00484en_HK
dc.identifier.authorityHuang, JD=rp00451en_HK
dc.description.naturelink_to_OA_fulltexten_US
dc.identifier.doi10.1074/jbc.C300328200en_HK
dc.identifier.pmid12917423-
dc.identifier.scopuseid_2-s2.0-0141960168en_HK
dc.identifier.hkuros91966-
dc.relation.referenceshttp://www.scopus.com/mlt/select.url?eid=2-s2.0-0141960168&selection=ref&src=s&origin=recordpageen_HK
dc.identifier.volume278en_HK
dc.identifier.issue41en_HK
dc.identifier.spage39578en_HK
dc.identifier.epage39582en_HK
dc.identifier.isiWOS:000185713800040-
dc.publisher.placeUnited Statesen_HK
dc.identifier.scopusauthoridTanner, JA=35513993000en_HK
dc.identifier.scopusauthoridWatt, RM=7102907536en_HK
dc.identifier.scopusauthoridChai, YB=55244155900en_HK
dc.identifier.scopusauthoridLu, LY=8686996700en_HK
dc.identifier.scopusauthoridLin, MC=7404816359en_HK
dc.identifier.scopusauthoridPeiris, JSM=7005486823en_HK
dc.identifier.scopusauthoridPoon, LLM=7005441747en_HK
dc.identifier.scopusauthoridKung, HF=7402514190en_HK
dc.identifier.scopusauthoridHuang, JD=8108660600en_HK

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