05-病毒学virus巴斯德研究所

ReplicationofDNAVirusesIPSTetsuyaToyodaFeb.28,20061.Replication2.TranscriptionDNARNAProteinDNApolymerasesDNAdepedentRNApolymerasesCentralDogmaDNARNAProteinPrionRNAvirusesViroidsSiRNARNAdependentRNApolymerasePrPscReversetranscriptaseRetrovirusesHBVRibozymeRibozymeRNAWorld:UniqueGeneExpressionlymphocytebrainheartlungkidneyspleenintestineInfectedmonocytePrimaryviremiaSecondaryviremiaSubcutanealhaemangitisSkinrushFreevirionsLocallymphnodeantibodiesSystemiclymphnodesSSPEvirusDropletinfectionKoplikspotLocalinfectionSystemicinfectionlypmocytepeniaReplicationofvirusinvivoMeaslesvirusmRNANS1PB1-F2adsorptionParentvirusendocytosisMembranefusionendosomeuncoatingnucleusHA、NA、M2Transcription,replicationtranslationM1、NS2buddingreleaseNewvirusReplicationofvirusintheinfectedcellInfluenzavirusStreamofgeneticinformationofDNAviruses＜Group1viruses＞＜Group2viruses＞＜Group3viruses＞＜Group4viruses＞＜Group5viruses＞＜Group7viruses＞dsDNA(+)mRNAdsDNA(+)ssDNA(+)dsRNA(+)ssRNA(+)ssRNA(-)ssRNA(-)ssDNA(-)ssRNA(+)dsDNA(+)mRNAStreamofgeneticinformationofRNAviruses＜Group6viruses＞ssRNA(+)ssRNAmRNAmRNA（±）ByBaltimoreTranscription,Replicationingeneralenzymecis-factortrans-factor(cis-element)Transcription:mRNAsynthesistranscriptasepromotertranscriptionfactorterminatorenhancer,silenceractivatorReplication:genomesynthesisreplicaseoriginreplicationfactorTranscription,Replicationofvirusesenzymecis-factortrans-factor(cis-element)Transcription:mRNAsynthesisRNApolIIpromotertranscriptionfactorterminatorenhancer,silenceractivatorReplication:genomesynthesisreplicaseoriginreplicationfactor(virus-encoded,pola)HostbasaltranscriptionfactorsCascadeofgeneexpressionofDNAvirusesEarlytranscriptionLatetranscriptionDNApolymeraseGenomereplicationStructuralproteinsInitiationofDNAreplicationProteinprimingProteinsthatactatreplicationforksProteinsatthereplicationforkProteinprimingOkazakifragmentDNAvirusreplicationReplicationcycleofvacciniavirusexceptionVacciniavirustranscriptionfactorsVacciniaviruspromotersequencesPoxvirusconcatemerresolutionsequenceVacciniavirusDNAreplicationTranscriptionmapofadenovirustype2exonintronSplicingofAdenovirusadenovirusDNAreplicationreplicationinitiatedonasingle-strandedDNAreplicationonduplextemplatesGenomicorganizationofSV40earlylateRegulatoryregionsofSV40oriCorepromotermotifsPreinitiation-complexTBP2BasalandactivatedtranscriptionGeneticanalysisofviruses1.Mutation2.Recombination3.Phenotypemixing(pseudotype)4.ComplementationApplicationofgeneticsMutationerrorReplicase:DNApolymerase1/109nucleotidesRNApolymerase1/103nucleotidesProofreadingactivity(exonucleaseactivity)SGKWVRELILYDWildtype(WT)gacgggaaatgggtgagagaactgattctgtatgacSGKWERELILYDPointmutationgacgggaaatgggagagagaactgattctgtatgacSGKWVRTDSVSTOPDeletiongacgggaaatgggtg**agaactgattctgtatgacSGKWVKRTDSVSTOPInsertiongacgggaaatgggtgaagagaactgattctgtatgacRecombinationRe-assortantSegmentedvirusPhenotypemixingPsuedvirustechnologyComplementationxxxxxxxxxxabortrescuetsmutantTemperaturesensitivemutant=pointmutationxxxxx39oC37oCxabortFunctionalproteinNon-functionalprotein