Investigations on the RNA binding and phosphorylation of groundnut bud necrosis virus nucleocapsid protein
Identifieur interne : 000425 ( Istex/Corpus ); précédent : 000424; suivant : 000426Investigations on the RNA binding and phosphorylation of groundnut bud necrosis virus nucleocapsid protein
Auteurs : Amruta S. Bhat ; H. S. SavithriSource :
- Archives of Virology [ 0304-8608 ] ; 2011-12-01.
Abstract
Abstract: Groundnut bud necrosis virus belongs to the genus Tospovirus, infects a wide range of crop plants and causes severe losses. To understand the role of the nucleocapsid protein in the viral life cycle, the protein was overexpressed in E. coli and purified by Ni-NTA chromatography. The purified N protein was well folded and was predominantly alpha-helical. Deletion analysis revealed that the C-terminal unfolded region of the N protein was involved in RNA binding. Furthermore, the N protein could be phosphorylated in vitro by Nicotiana benthamiana plant sap and by purified recombinant kinases such as protein kinase CK2 and calcium-dependent protein kinase. This is the first report of phoshphorylation of a nucleocapsid protein in the family Bunyaviridae. The possible implications of the present findings for the viral life cycle are discussed.
Url:
DOI: 10.1007/s00705-011-1110-0
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TocLevels="0"><ArticleID>1110</ArticleID><ArticleDOI>10.1007/s00705-011-1110-0</ArticleDOI><ArticleSequenceNumber>5</ArticleSequenceNumber><ArticleTitle Language="En" OutputMedium="All">Investigations on the RNA binding and phosphorylation of groundnut bud necrosis virus nucleocapsid protein</ArticleTitle><ArticleCategory>Original Article</ArticleCategory><ArticleFirstPage>2163</ArticleFirstPage><ArticleLastPage>2172</ArticleLastPage><ArticleHistory><RegistrationDate><Year>2011</Year><Month>9</Month><Day>6</Day></RegistrationDate><Received><Year>2011</Year><Month>4</Month><Day>17</Day></Received><Accepted><Year>2011</Year><Month>9</Month><Day>6</Day></Accepted><OnlineDate><Year>2011</Year><Month>9</Month><Day>24</Day></OnlineDate></ArticleHistory><ArticleCopyright><CopyrightHolderName>Springer-Verlag</CopyrightHolderName><CopyrightYear>2011</CopyrightYear></ArticleCopyright><ArticleGrants Type="Regular"><MetadataGrant Grant="OpenAccess"></MetadataGrant><AbstractGrant Grant="OpenAccess"></AbstractGrant><BodyPDFGrant Grant="Restricted"></BodyPDFGrant><BodyHTMLGrant Grant="Restricted"></BodyHTMLGrant><BibliographyGrant Grant="Restricted"></BibliographyGrant><ESMGrant Grant="Restricted"></ESMGrant></ArticleGrants></ArticleInfo><ArticleHeader><AuthorGroup><Author AffiliationIDS="Aff1"><AuthorName DisplayOrder="Western"><GivenName>Amruta</GivenName><GivenName>S.</GivenName><FamilyName>Bhat</FamilyName></AuthorName></Author><Author AffiliationIDS="Aff1" CorrespondingAffiliationID="Aff1"><AuthorName DisplayOrder="Western"><GivenName>H.</GivenName><GivenName>S.</GivenName><FamilyName>Savithri</FamilyName></AuthorName><Contact><Phone>+91-80-22932310</Phone><Fax>+91-80-23600814</Fax><Email>bchss@biochem.iisc.ernet.in</Email></Contact></Author><Affiliation ID="Aff1"><OrgDivision>Department of Biochemistry</OrgDivision><OrgName>Indian Institute of Science</OrgName><OrgAddress><City>Bangalore</City><Postcode>560012</Postcode><State>Karnataka State</State><Country Code="IN">India</Country></OrgAddress></Affiliation></AuthorGroup><Abstract ID="Abs1" Language="En" OutputMedium="All"><Heading>Abstract</Heading><Para>Groundnut bud necrosis virus belongs to the genus <Emphasis Type="Italic">Tospovirus</Emphasis>, infects a wide range of crop plants and causes severe losses. To understand the role of the nucleocapsid protein in the viral life cycle, the protein was overexpressed in <Emphasis Type="Italic">E. coli</Emphasis> and purified by Ni-NTA chromatography. The purified N protein was well folded and was predominantly alpha-helical. Deletion analysis revealed that the C-terminal unfolded region of the N protein was involved in RNA binding. Furthermore, the N protein could be phosphorylated <Emphasis Type="Italic">in vitro</Emphasis> by <Emphasis Type="Italic">Nicotiana benthamiana</Emphasis> plant sap and by purified recombinant kinases such as protein kinase CK2 and calcium-dependent protein kinase. This is the first report of phoshphorylation of a nucleocapsid protein in the family <Emphasis Type="Italic">Bunyaviridae</Emphasis>. The possible implications of the present findings for the viral life cycle are discussed.</Para></Abstract><AbbreviationGroup><Heading>Abbreviations</Heading><DefinitionList><DefinitionListEntry><Term>GBNV</Term><Description><Para>Groundnut bud necrosis virus</Para></Description></DefinitionListEntry><DefinitionListEntry><Term>TSWV</Term><Description><Para>Tomato spotted wilt virus</Para></Description></DefinitionListEntry><DefinitionListEntry><Term>RNP</Term><Description><Para>Ribonucleoprotein</Para></Description></DefinitionListEntry><DefinitionListEntry><Term>N protein</Term><Description><Para>Nucleocapsid protein</Para></Description></DefinitionListEntry><DefinitionListEntry><Term>CK2</Term><Description><Para>Protein kinase CK2</Para></Description></DefinitionListEntry><DefinitionListEntry><Term>CDPK</Term><Description><Para>Calcium-dependent protein kinase</Para></Description></DefinitionListEntry></DefinitionList></AbbreviationGroup><ArticleNote Type="ESMHint"><Heading>Electronic supplementary material</Heading><SimplePara>The online version of this article (doi:<ExternalRef><RefSource>10.1007/s00705-011-1110-0</RefSource><RefTarget Address="10.1007/s00705-011-1110-0" TargetType="DOI"></RefTarget></ExternalRef>) contains supplementary material, which is available to authorized users.</SimplePara></ArticleNote></ArticleHeader><NoBody></NoBody></Article></Issue></Volume></Journal></Publisher></istex:document></istex:metadataXml><mods version="3.6"><titleInfo lang="en"><title>Investigations on the RNA binding and phosphorylation of groundnut bud necrosis virus nucleocapsid protein</title></titleInfo><titleInfo type="alternative" contentType="CDATA"><title>Investigations on the RNA binding and phosphorylation of groundnut bud necrosis virus nucleocapsid protein</title></titleInfo><name type="personal"><namePart type="given">Amruta</namePart><namePart type="given">S.</namePart><namePart type="family">Bhat</namePart><affiliation>Department of Biochemistry, Indian Institute of Science, 560012, Bangalore, Karnataka State, India</affiliation><role><roleTerm type="text">author</roleTerm></role></name><name type="personal" displayLabel="corresp"><namePart type="given">H.</namePart><namePart type="given">S.</namePart><namePart type="family">Savithri</namePart><affiliation>Department of Biochemistry, Indian Institute of Science, 560012, Bangalore, Karnataka State, India</affiliation><affiliation>E-mail: bchss@biochem.iisc.ernet.in</affiliation><role><roleTerm type="text">author</roleTerm></role></name><typeOfResource>text</typeOfResource><genre type="research-article" displayLabel="OriginalPaper" authority="ISTEX" authorityURI="https://content-type.data.istex.fr" valueURI="https://content-type.data.istex.fr/ark:/67375/XTP-1JC4F85T-7">research-article</genre><originInfo><publisher>Springer Vienna</publisher><place><placeTerm type="text">Vienna</placeTerm></place><dateCreated encoding="w3cdtf">2011-04-17</dateCreated><dateIssued encoding="w3cdtf">2011-12-01</dateIssued><copyrightDate encoding="w3cdtf">2011</copyrightDate></originInfo><language><languageTerm type="code" authority="rfc3066">en</languageTerm><languageTerm type="code" authority="iso639-2b">eng</languageTerm></language><abstract lang="en">Abstract: Groundnut bud necrosis virus belongs to the genus Tospovirus, infects a wide range of crop plants and causes severe losses. To understand the role of the nucleocapsid protein in the viral life cycle, the protein was overexpressed in E. coli and purified by Ni-NTA chromatography. The purified N protein was well folded and was predominantly alpha-helical. Deletion analysis revealed that the C-terminal unfolded region of the N protein was involved in RNA binding. Furthermore, the N protein could be phosphorylated in vitro by Nicotiana benthamiana plant sap and by purified recombinant kinases such as protein kinase CK2 and calcium-dependent protein kinase. This is the first report of phoshphorylation of a nucleocapsid protein in the family Bunyaviridae. 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