细胞凋亡线粒体途径的调控

细胞凋亡线粒体途径的调控Regulationofmittochondrialapoptoticpathways高方远马欣荣康海岐ExecutionofmitochondrialapoptosissignalsReleaseofcytochromecReleaseofSmacReleaseofapoptosis-inducingfactorReleaseofendonucleaseGReleaseofcytochromecCytocApaf--1Cas-9Cas-3CleaveintracelluarsubstratesChr浓缩DNA分解核膜裂解等+dATP/ATPReleaseofSmacReleaseofapoptosis-inducingFactor(AIF)57KD黄素蛋白AIF(Mit)NucleusChr浓缩DNA降解IndependentofcaspaseactivationandoxidoreductasesactivityofAIFReleaseofendonucleaseGEndoGreleaseDNAfragmentapoptosisIndependentofcaspaseactivationBcl-2familyFeaturesofmitochondria-initiatedapoptosisMultiplefactorsfunctiontotriggercelldeathinconjunctionandinparallelways.thepathwayisabletofeed-forwardandamplifiedtheapoptoticsignal.Evenwhencaspase-dependentandcaspase-independentpathwaycannotfunctionproperly,mitochondrialdysfunctionmayleadtocelldeath.Thebestwaytopreventcelldeathistoblockapoptoticsignalsbeforemitochondrialdamageoccurs.Featuresofmitochondria-initiatedapoptosis1.Disruptionofelectrontransportandenergymetabolism.2.Releaseofcaspase-activatingproteins.3.Alterationofcellularreduction-oxidationpotentialThreegeneralmechanismsRegulationofmittochondrialapoptoticpathwaysRegulationofmitochondrialapoptoticsignals.Mitchondrialresponsetoapoptoticsignals.Lossofmitchondrialfunctionsduringapoptosis.RegulationofmitochondrialapoptoticsignalsTanslocationoftheBH3-onlyfamilyofproteinstomitochondrailCleavageofBidPhosphorylationofBadDisassaciationofBimTranscriptionalregulationofBH3-onlyproteinsTanslocationofotherproteinstomitochondraduringapoptosisTanslocationoftheBH3-onlyfamilyofproteinstomitochondrailOthercellularcompartmentsmitBH3-onlyproteins(Bid,Bad,Bim,Noxa,Puma)Bcl-2famillyCausemitdamageReleasecytoc,Smac,AIF,EndoGetc.CleavageofBidCellsurfacedeathreceptorsFas/CD95TNFR1DR-3,4TRAIL-R1,DR5TRAIL-R2FasLTNFApo3LApo2L/TRAILReceptorsligandextracytoplasmicCys-richDeathdomaintailDISC(死亡诱导信号复合体)Death-signalingpathwayTypeIcellTypeIIcell（肝细胞）Cas-8Bcl-2apoptosisBidtBidAmlifiedsignalsenoughNotenoughcleavetranslocationCleavageofBidmitmitmitBid(cytocolicInlivingcells)tBid(cyto)tBid(mit)CytoCrealeaseDomain:helicese4-6Specificity:mit-specificlipid,casdiolipin(心磷脂)Efficiency:enhancedbymyristorylation(豆蔻酰化)ofN-terminalglycineresidueoftBidexposedaftercas-8cleavageCas-8cleavesignalsFas/FasLapoptosisOtherregulationpathwayOthercas(cas-3)cas-8Andprotases(粒酶B,Bid(cyto)otherBH3-onlyproteins溶原体蛋白酶)(inactiveBad)tBidactiveBadtBid(mit)reaseapoptosisPhosphorylationofBadapoptosisBad(Incytosol)14-3-3proBcl-2(active)inactive-Survivalsignals+trophicfactors(Incytosol)(Inmit)Akt,PKAThecriticalsiteofphosphrylationBysurvivalfactorsBH3domainofBadSer112Ser136Ser155apoptosisSer155kinasesElectrostatic（静电）and空间限制Ser112136155Bcl-xl14-3-3p70S6kinase纳巴霉素Phosphatases(Invitro)Bad-pdephosphorulatedBadCalcineurin(钙调磷酸酶)Proteinphosphatase1aProteinphosphatase2AProteinphosphatase1aisaRas-activatedBadphosphatasethatregulatesinterleukin-2deprivation-inducedapoptosisEMBOJ.19:2237-2246Materials:interleukin-2(IL-2)-dependentmurineT-celllineMethods:Usingtheyeasttwo-hybridsystem,glutathioneS-transferase(GST)fusionproteinsandco-immunoprecipitationtechniques,Results:Badinteractswithproteinphosphatase1a(PP1a).Bad+IL-2ActivatedkinasesAkt,Psk,PKAetc.Badphosphorylation-IL-2PP1aRasBaddephosphorylationAkadaicacid(冈田酸)apoptosisHowthesephosphatasesareregulatedinvivobyapoptoticsignalsremainstobeinvested??InteractionofBadandPP1phosphatase.InteractionofBadandthecatalyticsubunitofPP1phosphataseinthetwo-hybridsystem.DisassaciationofBimmitcytosolBimMicrotubulecomplexesLC8Bim-deficientlymphocytesareresistanttocertainapoptoticstimuli.Cytokinedeprivation（细胞因子缺失）Calciumionflux（钙离子流出）Microtubulepertubation(微管干扰)AbundanceofBimBeregulatedattheleveloftranscription?TranscriptionalregulationofBH3-onlyproteinsApoptosisrequiresnewproteinssynthesistranscriptionalregulationofBH3-onlyproteinmaybeimportantforapoptosisnewlygeneratedproteinmitDirectlytargetAKTFKHR-L1(叉头转录因子)BimtranscriptionNeuronandhematopoieticprogenitors(造血干细胞)-Cytokine显性失活的C-JunHRKtranscription①②-NGF（Inneurons)③④apoptosisX-irradiationDNAdamageP53Noxa,Puma(BH3-onlymember)TranscriptionalregulationofBH3-onlyproteinplayamajorroleinDNAdamageapoptosisBH3domaintriggerCytochromecreleaseCaspasesactivationNoxa,aBH3-OnlyMemberoftheBcl-2FamilyandCandidateMediatorofp53-InducedApoptosis2000,science288:1053TanslocationofotherproteinstomitochondraduringapoptosismitP53TR3P53AIPLKB1MitdamageapoptosisCytocetc.????.Targetingp53tomitochondriaofp53-deficientcellsissufficienttoinduceapoptosis.Roleofmitochondrialp53inregulatingapoptosis.CytochromecReleaseandApoptosisInducedbyMitochondrialTargetingofNuclearOrphanReceptorTR32000，science289:1159-1164.MitochondrialtargetingofTR3regulatesmitochondrialactivity.(A)TR3expressionisrequiredforcytochromecreleaseinresponsetoapoptosisinducers.(B)MitochondrialtargetingofTR3isassociatedwithcytochromecrelease.(C)TR3/1blockscytoplasmiclocalizationofTR3.(D)TR3/1inhibitsMM11453-inducedapoptosis.Quantificationofapoptoticcellsin400TR3/1-transfectedornontran