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NSs Encoded by Groundnut Bud Necrosis Virus Is a Bifunctional Enzyme 英文参考文献
NSsEncodedbyGroundnutBudNecrosisVirusIsa
BifunctionalEnzyme
BhushanLokesh1,PanigrahiR.Rashmi2,BhatS.Amruta1,DharmaiahSrisathiyanarayanan1,MathurR.N.
Murthy2,HandanahalS.Savithri1*
1DepartmentofBiochemistry,IndianInstituteofScience,Bangalore,Karnataka,India,2MolecularBiophysicsUnit,IndianInstituteofScience,Bangalore,Karnataka,India
Abstract
Groundnutbudnecrosisvirus(GBNV),amemberofgenusTospovirusinthefamilyBunyaviridae,infectsalargenumberof
leguminosaeandsolanaceaeplantsinIndia.Withaviewtoelucidatethefunctionofnonstructuralprotein,NSsencodedby
thesmallRNAgenome(SRNA),theNSsproteinofGBNV-tomato(Karnataka)[1]wasover-expressedinE.coliandpurified
byNi-NTAchromatography.ThepurifiedrNSsproteinexhibitedanRNAstimulatedNTPaseactivity.Further,thisactivitywas
metal ion dependent and was inhibited by adenosine 59 (b, c imido) triphosphate, an ATP analog. The rNSs could also
hydrolyzedATP.Interestingly,inadditiontotheNTPaseanddATPaseactivities,therNSsexhibitedATPindependent59RNA/
DNA phosphatase activity that was completely inhibited by AMP. The 59 a phosphate could be removed from ssDNA,
ssRNA,dsDNAanddsRNAthusconfirmingthatrNSshasanovel59aphosphataseactivity.K189AmutationintheWalker
motifA(GxxxxGKT)resulted incompletelossofATPaseactivity,butthe59phosphataseactivitywasunaffected.Onthe
other hand, D159A mutation in the Walker motif B (DExx) resulted in partial loss of both the activities. These results
demonstrateforthefirsttimethatNSsisabifunctionalenzyme,whichcouldparticipateinviralmovement,replicationorin
suppressionofthehostdefensemechanism.
Citation: LokeshB,RashmiPR,AmrutaBS,Srisathiyanarayanan D,MurthyMRN,etal.(2010)NSsEncodedbyGroundnutBudNecrosis VirusIsaBifunctional
Enzyme.PLoSONE5(3):e9757.doi:10.1371/journal.pone.0009757
Editor:PeterSommer,InstitutPasteurKorea,RepublicofKorea
ReceivedNovember12,2009;AcceptedFebruary22,2010;PublishedMarch18,2010
Copyright: ? 2010 Lokesh et al. This is an open-access article distributed under the terms of the Creative Common
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