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1、JBCPapersinPress.PublishedonOctober5,2012asManuscriptR112.416925Thelatestversionisathttp://www.jbc.org/cgi/doi/10.1074/jbc.R112.416925THERESTRICTIONFACTORSOFHUMANIMMUNODEFICIENCYVIRUS112ReubenS.Harris,JuddF.HultquistandDavidT.Evans1DepartmentofBiochemistry,Mo

2、lecularBiologyandBiophysics,DepartmentofGenetics,CellBiologyandDevelopment,InstituteforMolecularVirologyUniversityofMinnesota,Minneapolis,MN,554552DepartmentofMicrobiologyandImmunobiology,HarvardMedicalSchool,NewEnglandPrimateResearchCenter,OnePineHillDrive,S

3、outhborough,MA01772-9102Addresscorrespondenceto:ReubenS.Harris,321ChurchStreetS.E.,6-155JacksonHall,Minneapolis,MN55455.E-mail:rsh@umn.eduSUMMARYvirusrecoveredinthecellculturemediumCellularproteinscalled‘restrictionafter1to2daysofincubation.SuchassaysDownload

4、edfromfactors’canserveaspowerfulblockadesareideallydoneoverarangeofrestrictiontoHIVreplication,buttheviruspossessesfactorexpression,withthehighestlevelselaboratestrategiestocircumventtheseoftenimposinglog-scaledropsinviralbarriers.First,wediscussgeneralinfect

5、ivity(Fig.1B).This‘singlecycle’hallmarksofarestrictionfactor.Then,assayforvirusreplicationisusefulforhttp://www.jbc.org/wereviewhowtheviralVifproteintestingtheimpactofviralmutationsand/orprotectstheviralgenomefromlethalrestrictionfactorvariations.levelsofcDNA

6、deaminationbypromotingSecond,ifarestrictionfactorisaAPOBEC3proteindegradation,howthetruethreattoviralreplication,thenthebyguestonAugust12,2018viralVpu,Env,andNefproteinsfacilitatepredecessorsofHIVinvariablyevolvedaninternalizationanddegradationoftheequallypot

7、entcounter-restrictionvirus-tetheringproteinTetherin/BST-2,mechanismthatstillexistsinthepresentdayandhowtheviralVpxproteinpreventsvirus.Forinstance,titratingacounter-theprematureterminationofreverserestrictionfactorintotheaforementionedtranscriptionbydegradin

8、gthedNTPasesinglecycleinfectivityexperimentcanresultSAMHD1.TheseHIVrestrictionandinafullrecoveryofviralinfectivitydespitecounter-restrictionmechanismssuggestthepresenceofanactiverestricti