分子細(xì)胞生物學(xué)進(jìn)展：細(xì)胞信號(hào)轉(zhuǎn)導(dǎo)

細(xì)胞信號(hào)轉(zhuǎn)導(dǎo)CellSignalingSignalTransduction分子細(xì)胞生物學(xué)的核心問(wèn)題GeneProteinBiologicalprocess細(xì)胞外信號(hào)分子受體蛋白細(xì)胞內(nèi)信號(hào)轉(zhuǎn)導(dǎo)分子效應(yīng)蛋白細(xì)胞反應(yīng)細(xì)胞信號(hào)通路的基本組成生命體是一架精密的機(jī)器，但她有比機(jī)器高一層次的調(diào)節(jié)程序“有機(jī)體”、“穩(wěn)態(tài)”、“適應(yīng)性反應(yīng)”……任何生物學(xué)現(xiàn)象都有復(fù)雜的調(diào)控過(guò)程ApoptoticSignalingSignalingNetworksignalreceptorSignaltransductionpathwayGeneexpression門(mén)控離子通道酶偶聯(lián)受體G偶聯(lián)受體核受體信號(hào)通路（signalingpathwayorsignaltransductionpathway）Jak-Statpathway：Stat是轉(zhuǎn)錄因子。在激活后，從細(xì)胞質(zhì)轉(zhuǎn)位到細(xì)胞核內(nèi)，調(diào)控基因表達(dá)。信號(hào)通路（signalingpathwayorsignaltransductionpathway）MAPKpathway:(cascademodule級(jí)聯(lián)式方式)信號(hào)通路（signalingpathwayorsignaltransductionpathway）GPCR-cAMPpathway:第二信使cAMP核受體(Nuclearreceptor)轉(zhuǎn)錄因子(Transcriptionfactor)Hormone(ligand)皮質(zhì)醇信號(hào)轉(zhuǎn)導(dǎo)通路的分子組成Signalingcomponentsaremainlyproteins：1.Enzymes：RTK,adenylatecyclase,phospholipase,kinase，specificationandamplifer(特異性和信號(hào)放大)。2.Protein-proteininteraction:bindingdomain(結(jié)合域)，specification(特異性)。3.Proteinmodification：active/inactive(激活/失活)、interaction(蛋白質(zhì)相互作用)第二信使（Secondmessenger)：在傳遞過(guò)程中產(chǎn)生的小分子：cAMP、cGMP、IP3、DG等，放大信號(hào)。

細(xì)胞信號(hào)轉(zhuǎn)導(dǎo)的基本特點(diǎn)

Specificity(高度專(zhuān)一性)；Amplification(信號(hào)放大)；TransientandDynamic(一過(guò)性、或可逆性、或分子開(kāi)關(guān))；Amolecularswitchisamoleculethatcanbereversiblyshiftedbetweentwoormorestablestates.Integration(信號(hào)整合)；Crosstalkandnetwork（信號(hào)通路的相互聯(lián)系）細(xì)胞信號(hào)轉(zhuǎn)導(dǎo)的調(diào)節(jié)

信號(hào)轉(zhuǎn)導(dǎo)的動(dòng)態(tài)性（dynamic）表明信號(hào)轉(zhuǎn)導(dǎo)是被調(diào)控的。信號(hào)轉(zhuǎn)導(dǎo)分子是調(diào)控的靶標(biāo)(signaling

componentsarecontrolled)誘導(dǎo)(induction)：自身通路或其它途徑；正反饋環(huán)；…

保持(extension)：活性保持的長(zhǎng)短是被控制的；下調(diào)(down-regulation)：一過(guò)性：

分子與分子的解離(disassociation)，分子降解(degradation)，與調(diào)節(jié)蛋白結(jié)合(bindingwithregulator)，蛋白質(zhì)修飾的可逆性(reversedproteinmodification)，負(fù)反饋(negativefeedbackloop)SENP1HypoxiaHIF1asumoHIF1aEffectsPIASysumosumoHIF1aSUMOylationinhypoxiasignaling012346(hr)hypoxiaSUMO-HIF1a024610(hr)hypoxiaSENP1G3PDHRT-PCRInducecomponentexpression誘導(dǎo)信號(hào)分子表達(dá)Dynamicsignaling信號(hào)轉(zhuǎn)導(dǎo)的動(dòng)態(tài)性ReversedSUMOylation蛋白質(zhì)修飾的可逆性Bindingwithregulator蛋白質(zhì)相互結(jié)合Feedbackloop反饋環(huán)HIF1ahypoxiaSENP1VEGFVEGFVEGFsignallingEndothelialcellR2SENP1regulateshypoxiasignalingthroughpositivelyfeedbackloopsi-NSsi-SENP1Protein-ProteinInteractionBasisofcellsignalingdynamic,specificity,amplification,……MediatedbyproteinmodificationmostlyProteinpost-translationmodification

>200kinds

Phosphorylation,Ubiquitination,SUMOylation,…SignalTransductionpathwayEffectProtein-proteininteractionProteinmodificationRegulationCellSignalingandMolecularBasisSignalEffectSignalTransductionpathwayEffect?Transductionpathway?

Signal?TransductionpathwayEffect1:SignalFunctionalscreeningstrategyReadoutSignalOverexpressionsiRNA,anti-senes,dominantmutant,SpecificinhibitorBiochemistryapproaches…GeneexpressionSpecificactivityEffectTransductionpathway?Transductionpathway?TransductionpathwayEffect?2:SignalSignalOverexpression/transgenicmousesiRNA,anti-senes,dominantmutant/knockoutmouseSpecificinhibitor,…inAnimalmodel,Cell,molecular,TransductionpathwayEffect?ProteinSUMOylationandsignalingUbiquitin泛素SUMO類(lèi)泛素2004NobelPrizeinChemistry--Ubiquitin-mediatedproteindegradationAaronCiechanoverAvramHershkoIrwinRoseG-G(SENPs)E1+E2+E3+SUMOtargetSUMOtargetSUMOSUMO-SpecificProteasesProteinSUMOylationSUMO:expressedinalleukaryotes(yeast,plantsandanimals).TheroleofSUMOylation:fromthestandpointofbiochemistryNormalRanGAP1MutatedRanGAP1SomefactsaboutSUMOylationLocalizationInteractionRegulationofgeneexpressionNatureReviews-MolCellBiol8:947,2007SUMOylatedproteinsarefoundthroughoutthecell“SUMOmotif:yKxE”SUMOylationSystemE1:1E2:1E3:15ormoreSENP:6TargetTargetsusuE1+E2suSENP+E3SENP1SENP2SENP3SENP5SENP6SENP7HDQCSentrin/SUMO-SpecificProtease(SENP)SENPsTargets/Functions?Regulators?ResearchFocus:Twostrategies:Functionalscreening

GeneknockoutFunctionalscreeningstrategySENPs?Readout(targets)StimuliAndrogenreceptor（AR）andco-factorsareSUMOylatedproteinsARSnSnp300SRC-1,GRIP1SnSnHDAC1AREARARAndrogenCytoplasmNucleusSENP1SENP1mSENP2SENP2mSENP3SENP3mSENP6SENP6mSENP7aSENP7amSENP7bSENP7bmSENP8SENP8mSUMO-1SUMO-2basalR1881ContrLuciferaseActivitySENP1specificallyenhancesARactivityARARELuciferaseASENP1andARsignalingARSnSnp300SRC-1,GRIP1SnSnHDAC1AREARARAndrogenCytoplasmNucleusSENP1MCB.2005Vector00.5123Chase(hr)AS35-SENP1RA1881+BicMockBpCMV-GFP-SENP1-LNCaPcellsAndrogeninducesSENP1expression&stabilizationR1881E1ARSENP1promoterAREAndrogenCJBC,2007SENP1regulatesARsignalingthroughpositivefeedbackloopSENP1ARAndrogenProstateCancerInductionofSENP1expressioninprostatecancercells

HGPINPCNeoplasia,2006Twostrategies:Over-expressioninnormalcellsorcellswithlowerexpressionlevelDown-regulationincellswithhigherexpressionlevelSENP1(tg)ControlPIN

formationinSENP1transgenicmiceSENP1高表達(dá)能誘導(dǎo)LNCaP細(xì)胞形成癌細(xì)胞轉(zhuǎn)移樣特征EMT(Epithelial-MesenchymalTransition)VectorSENP1SENP1mutant正常前列腺上皮細(xì)胞癌性增生PIN原位癌轉(zhuǎn)移癌SENP1SENP1SENP1其他因子?PopulationDoublingsThedecreaseincellgrowthofRas-transformedSENP1-/-MEFcellsPassages12345+/+-/-VectorRasb-galstainingIncreasedoncogenic-inducedsenescenceinSENP1-/-MEFcells癌基因+SENP1細(xì)胞衰老-SENP1細(xì)胞衰老腫瘤生長(zhǎng)抑制SENP1在細(xì)胞轉(zhuǎn)化中的作用SENP1在前列腺癌發(fā)生中的可能作用正常前列腺上皮細(xì)胞癌性增生PIN原位癌轉(zhuǎn)移癌WTSENP1TgVectorSENP1GeneknockoutstrategyGenemutationPhenotypesTargetsDayObservedratioExpectedratioE12.518/7519/75E13.519(14*)/14536/145E14.55(2)/4311/43E15.56(12)/9524/95>E16.52(2)/8822/88Postnatal0/36892/368AB+/+-/-n=4n=9n=5+/++/--/-RedBloodCells(relativelevels)CEmbryoniclethalityofSENP1-/-duetoanemia*Rednumberrefertodeadembryos-/-TUNELHSCBFU-eCFU-eRBCTer-119+ErythroblastAnemiaisduetoerythroblastapoptosisinSENP1-/-embryoFACS/Colony+/+ProteasomaldegradationPHDHIF1aubnVHLVHLHypoxiaSENP1HIF1aHIF1bNucleusCytoplasmsusuEpo,VEGF,Glut-1ProteasomaldegradationsuOHHIF1aHIF1aubnHIF1aOHHIF1aHIF1aSENP1regulatesHIF1aactivityunderhypoxiaCell,131:584,2007SENP1HypoxiaHIF1asumoHIF1aEffectsPIASysumosumoDynamicSUMOylationofHIF1aduringhypoxia012346(hr)hypoxiaSUMO-HIF1a024610(hr)hypoxiaSENP1G3PDHRT-PCRHypoxiainducesSENP1expressionthroughHIF1aHypoxiaHIF1aSENP1SENP1promoterSENP1HIF1aHypoxiaVEGFAngiogenesisSENP1regulateshypoxiasignalingthroughpositivefeedbackloopSENP1RegulatesAngiogenesissi-NSsi-SENP1Xuetal.JBiolChem,2010HIF1ahypoxiaSENP1VEGFVEGFR2VEGFsignalingSENP1inTumorAngiogenesisaSENP1aVEGFaCD31Case#1#2Zuoetal.UnpublishedDataSENP1andtumorangiogenesisSENP1HIF1aVEGFTumorAngiogenesishypoxiaSENP1+/+SENP1-/-c-kit+/+-/-EpoPCRcycleSENP1CD68SENP1CD45aSENP1SENP1ExpressesinBoneMarrow-DerivedCellsinTumorStro