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Download Student Worksheet Hands-on Activity Viral DNA Integration
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Hands-on Activity Viral DNA Integration Student Worksheet INTRODUCTION Thehumanimmunodeficiencyvirus(HIV)infectsanddestroyscellsoftheimmunesystemcalledhelper Tcells.Overtime,HIVinfectionweakensaperson’sabilitytofightotherinfectionsandsomediseases. TheadvancedstageofHIVinfectionistermedacquiredimmunodeficiencysyndrome(AIDS).An individualwithAIDShasaseverelyimpairedimmunesystem.AlthoughthereisnocureforAIDS,HIV infectioncanbecontrolledwithpropertreatmentandearlymedicalcare. HIVisaretrovirus.Likeallviruses,retrovirusescanonlyreplicatewithinhostcells.Theyusethehost cell’smachinerytomakecopiesoftheviralgenomeandproduceitsproteins.Retrovirusesare distinguishedfromothertypesofvirusesbytwokeystepsinthereplicationcycle:reversetranscription andintegration.AfterHIVinfectsahostcell,reversetranscriptionresultsintheproductionofadoublestrandedDNAcopyofthesingle-strandedHIVRNAgenome.Integrationthenresultsintheinsertionof thedouble-strandedDNAcopyintothehost’sgenome.ThehostcellthentreatstheintegratedHIVDNA aspartofitsowngenome. Inthisactivity,youwillmodeltheintegrationprocess. MATERIALS Forthisactivity,youwillneedpopbeadsofthreedifferentcolorstorepresent: • thedouble-strandedDNAgenomeofthehostcell(about40to50beads) • thesingle-strandedRNAgenomeofthevirus(about5to10beads) • thedouble-strandedDNAcopyofthevirus’sgenome(doublethenumberofbeadsusedforthe RNA) PROCEDURE 1.WatchananimationoftheHIVlifecycle(http://www.hhmi.org/biointeractive/hiv-life-cycle).Pay particularattentiontothesectiononreversetranscriptionandintegration.Takenotesontheenzymes involvedinthesetwosteps.YoumayhavenoticedintheanimationthateachHIVparticlecontainstwo copiesoftheHIVgenome.Eachcopyisasingle-strandedRNAmolecule. 2.GothroughtheHHMI“Click&Learn”activityentitled“VirusExplorer” (https://www.hhmi.org/biointeractive/virus-explorer).Inparticular,readthedefinitionsforgenome, structure,andenvelopebyclickingonthe“i”iconnexttoeachword.ClickonHIVandlookatboththe 3-Dmodelandcross-sectionimages.Thenanswerquestions#1-5below. www.BioInteractive.org February3,2017 Page1of4 Hands-on Activity Viral DNA Integration Student Worksheet QUESTIONS 1.DrawadiagramandexplainthebasiccharacteristicsofHIV.(Ifyouarefillingouttheformonyour computer,takeaphotoofyourdrawinganduploaditinthespaceprovided.) LabeledDiagram ExplanationofLabeledStructures 2.Definetheterm“genome”anddescribetheHIVgenome. 3.WhatisthehostcellforHIV?Howarethecompositionandsizeofthehostcellgenomedifferent fromthoseofHIV? 4.Whatistheenzymeresponsibleforreversetranscription?Isitaviralorhostenzyme? 5.WhatistheenzymeresponsiblefortheinsertionoftheviralDNAintothehostgenome?Isitaviralor hostenzyme? PROCEDURE 6.Youwillnowmakeamodelofthehostcellgenomeandvirusgenomeusingpopbeads.Usethe longerstrandtorepresenttheDNAofthehostcell.DrawadiagramofwhatyouconstructedinTable1 below. www.BioInteractive.org February3,2017 Page2of4 Hands-on Activity Viral DNA Integration Student Worksheet 7.Onceinsidethehostcell,anHIVenzymeproducesadouble-strandedDNAcopyoftheviralgenome. Usethethirdcolorofpopbeadstomodelthedouble-strandedDNAcopyoftheviralRNA.Drawa diagramofyourpop-beadmodelinthetablebelow.(Youcanalsotakephotosofyourmodelsand uploadthephotosinthespacesprovided.) Table1.ModelsofHostCellGenome,HIVGenome,andReverse-TranscribedViralDNA. Model Drawa diagramofthe modelyou constructed HostCellGenome HIVgenome DNAcopyofHIVgenome Characteristics (RNAorDNA; single-or doublestranded; relativesize) 8.Comparethethreemoleculesyouconstructed.Whatarethekeysimilaritiesanddifferences? www.BioInteractive.org February3,2017 Page3of4 Hands-on Activity Viral DNA Integration Student Worksheet 9.AftertheviralDNAisproduced,thenextstepintheHIVlifecycleistheinsertionofviralDNAintothe hostcellgenome.Usethedouble-strandedviralDNAandthehostDNApop-beadmodelstoconstructa modelshowinghowthevirus’sgenomeisintegratedintothehostgenome.Drawadiagramofyour pop-beadconstructionbelowandmakesureyoulabelyourdiagram.(Youcanalsotakeaphotoofyour modelanduploaditinthespaceprovided.) 10.Describethestepsinvolvedintheintegrationprocess.(Thinkaboutwhatyouhadtodowiththe beads.) 11.DescribewhathappenstotheviralDNAnowthatitisintegratedintothehostcellgenome. www.BioInteractive.org February3,2017 Page4of4