RNA结合蛋白与转录后调控.pptx

RNA结合蛋白与转录后调控RegulatoryfactorsformRNAdecayandtranslationRNAbindingproteins(RBPs)AUF1EffectofARE-BPsonthestabilityandtranslationofARE-containingmRNAsARE-BPsmRNAstabilityProteinexpressionTranslationalefficiencyAbundanceIncreaseDecreaseIncreaseDecreaseUpregulatedDownregulatedAUF1c-myc(42)c-myc(46)GM–CSF(55)c-fos(42,67)c-fos(53)IL-3(55)PTH(56)p21(48)GM–CSF(42)CyclinD1(48)TNF-alpha(42)GM–CSF(53,54)IL-3(55)HuRc-fos(59,63,67)p53(99,137)TNF-alpha(139)p21(69)TNF-alpha(139)MyoD(68)Cox-2(139)CyclinA(70)p21(48,68,69)CyclinB1(70)CyclinA(70)NOSII/iNOS(64)CyclinB1(70)GM–CSF(55)CyclinD1(48)Cox-2(71,173)NOSII/iNOS(64)IL-3(55)GM–CSF(59)VEGF(173)TNF-alpha(65,74,139)p53(99,137)Cox-2(71,139)IL-3(55,66)VEGF(62)Myogenin(68)Hel-N1TNF-alpha(74)NF-M(73)NF-M(73)GLUT1(72)GLUT1(72)GLUT1(72)HuDGAP-43(75–77)GAP-43(75,76)TTPc-fos(90)GM–CSF(81)GM–CSF(18,81,83–85,91)TNF-alpha(80)TNF-alpha(18,81,83–86,89,90)IL-2(82)Cox-2(87)IL-3(88)IL-2(82,90)IL-3(18,66,83,84,88)BRF1TNF-alpha(89,93)GM–CSF(55)IL-3(55,92,93)IL-3(55)TIA-1TNF-alpha(120)TNF-alpha(120)Cox-2(121)Cox-2(121)KSRPc-fos(90,93)NOSII/iNOS(102)NOSII/iNOS(102)TNF-alpha(90,93)IL-2(90,93)c-jun(93)CUG-BP2Cox-2(150)Cox-2(150)Cox-2(150)Nucleolinbcl-2(175)TINObcl-2(176)PAIP2VEGF(177)VEGF(177)AU-richElements(AREs)RNA-蛋白质相互作用(结合)的特点"Tet-on"systemisactivatedinthepresenceofdoxycyclinemRNA翻译研究特用技术新生蛋白分析。Polysome分离，PolysomalRNA,Polysomal蛋白质分离。其它经典技术。HuRRegulatesp21mRNAStabilizationbyUVLightWang,etal;MolCellBiol.2000，20(3):760–769.A。复合物形成在UVC辐射半小时后明显被诱导，与P21被诱导相吻合。B。竞争抑制试验说明复合物的特异性，Cold探针可竞争抑制复合物形成。C。EMSA后，UV交联，SDS-PAGE分离复合物，发现复合物中一条大约40Kd的复合物（单一蛋白与大约10个碱基的短片断转录物形成），说明有一35-40KdRNA结合蛋白被UVC诱导并与P213’-UTR结合。D。UVC诱导该未知复合物的趋势与P21被诱导一致。HuR表达降低后，p213′UTR与细胞浆蛋白间形成的复合物（在UVC辐射后）降低，p21mRNA稳定性降低（半衰期缩短），表达降低。DecreasedHuRexpressionlowersbindingtothep213′UTRandreducesp21mRNAstabilityandp21inductionbyUVC.(A)WesternblotanalysisofHuRexpressioninRKOcells,eitheruntransfected(untr.)ortransfectedwithpZeoSV2(−)HuR,expressingASHuR.Chosenclonalisolatesareshown.Blotsweresequentiallystripp