简介:Changesinthedistributionof1P1-antigeninthedevelopingchickretinahavebeenexaminedbyindriectimmunofluorescencestainingtechniqueusingthenovelmonoclonalantibody(MAb)1P1.Expressionofthe1P1antigenwasfoundtoberegulatedinradialaswellasintangentialdimensionoftheretina,beingpreferentiallyorexclusivelylocatedintheinnerandouterplexiformlayersoftheneuralretinadependingonthestagesofdevelopment,Withtheonsetoftheformationoftheinnerplexiformlayer1P1antigenbecomesexpressedintheretina.Withprogressingdifferentiationoftheinnerplexiformlayer1P1immunofluorescencerevealed2subbandsatE9and6subandsatE18,Atpostnatalstages(afterP3)immunoreactivitywasreducedinaninside-outsidesequenceleadingtothecompleteabsenceofthe1P1antigeninadulthood.1P1antigenexpressionintheouterplexiformlayerwasalsosubjecttodevelopmentalregulation.Thespation-temporalpatternof1P1antigenexpressionwascorrelatedwiththetimecourseofhistologicaldifferentationofchickretina,namelythesynapserichplexiformlayers.Whetherthe1P1antigenwasfunctionallyinvolvedindendriteextensionandsynapseformationwasdiscussed.
简介:在Saccharomycescerevisiae,必要基因CDC13编码telomeric遗传上并且身体上与Stn1p和Ten1p交往的搁浅单人赛的DNA有约束力的蛋白质,并且为telomere结束保护和telomere长度控制被要求。Ten1由参予telomere长度规定和染色体结束保护的分子的机制留下逃犯。在这个工作,我们用净化的recombinantCdc13p和Ten1p在胶化过滤分析观察了Cdc13p和Ten1p的一个弱相互作用。Ten1p本身展出一项弱DNA有约束力的活动,但是提高telomericTG1鈥吗?Cdc13p的DNA有约束力的能力。Cdc13p是有Ten1p的co-immunoprecipitated。在变异的ten1-55或ten1-66房间,在Ten1p和Cdc13p之间的损害相互作用与telomericDNA导致长得多的telomeres,以及Cdc13p的一个减少的协会。一致地,Ten1-55和Ten1-66异种蛋白质没能刺激telomeric在vitro的Cdc13p的DNA有约束力的活动。这些结果建议Ten1p提高telomericCdc13p到的DNA有约束力的活动否定地调整telomere长度。
简介:WereportedinthismanuscriptthatTGF-β1inducesapoptosisinAML12murinehepatocytes,whichisassociatedwiththeactivationofp38MAPKsignalingpathway.SB202190,aspecificinhibitorofp38MAPK,stronglyinhibitedtheTGF-β1-inducedapoptosisandPAI-1promoteractivity.TreatmentofcellswithTGF-β1activatesp38.Furthermore,over-expressionofdominantnegativemutantp38alsoreducedtheTGF-β1-inducedapoptosis.Thedataindicatethattheactivationofp38isinvolvedinTGF-β1-mediatedgeneexpressionandapoptosis.
简介:死亡联系域的蛋白质Daxx施加包括调停的许多报导功能经由激活c6月N终端从FasL发信号到apoptosisapoptosis的kinase(JNK),正式就职和抑制,和染色质改变的规定。它原来从酵母被克隆用船边交货对相似物体之连续感觉而形成心像口的细胞内部的尾巴的二混血儿的屏幕诱饵。而许多起始的报告仍然保持争论,Daxx在一系列压力信号包括UVirradiation,过氧化氢治疗和TGF-β治疗触发的apoptosis的规定起重要作用,是清楚的。在这评论,我们在Axinbeing上集中连接Daxx到p53的一个拴住的因素。
简介:CCAAT/enhancer有约束力的蛋白质α(C/EBPα)是transcriptional禁止细胞增殖的规章的因素,和其他的翻译开始生产二多肽,C/EBPαp30并且C/EBPαp42。由表示介绍,C/EBPαp30specifically禁止了基因的一个唯一的集合,这被揭示,包括MPP11,p84N5和SMYD2,它没被C/EBPαp影响42在两根QSG-7701hepatocyte房间线和QGY-7703hepatoma,cells.Semi量的RT-PCR分析独立地证实了这些结果。Chromatinimmunoprecipitation试金显示出30比C/EBPαp更强烈绑了在这些基因的倡导者的那C/EBPαp42。在C/EBPα是调整的down的临床的hepatoma在样品,所有三基因明确地由30在上面的C/EBPαp禁止了调整。然而,MPP11,p84N5和SMYD2genes的压抑可能直接不涉及C/EBPαp30-mediated生长抑制。我们的数据建议30调整的那C/EBPαp目标基因的一个唯一的集合并且多于C/EBPαp的dominant-negativeregulator42。
简介:Amurinemacrophage-likecelllineJ774,acquired,inresponsetoLPS,anabilitytokilltumornecrosisfactor(TNF)-insensitivetargetP815mastocytomacellswhereasanothercellline,P388D1didnot,LPStriggeredsignalingmechanismsbetweenthetwocelllineswerecomparedwithanaimtoinquireaboutthepossiblenatureoftheabove-mentioneddifference,TheresultswhowedthattwocelllinesrespondtoLPS-treatmentbyparallelactivationofbothphospholipasesCandA2(PLCandPLA2)toapproximatelythesameextent.ThemaximumresponseoftothenzymesofJ774cellswasnotedwithin10minthetreatmentwhereasthatofP388D1cellsrequiredmorethan20min,TheotherpropertiesofLPS-responsiveenzymesstudiedweresimilarbetweentwocelllines,includingActivationofPLCandPLA2andPKCinmacrophagesbyLPS.Ca2+augmentationofenzymeactivation,participationofguaninenucleotidebinding(G)proteinsintheinitialactivationpreocesses,andinhibitionofenzymeactivationbythepriortreatmentofcellswithcholeraorpertussistoxinsetc.Moreover,LPS-triggeredactivationofPLCandPLA2wasfoundtobefollowedbytheincreaseofPKCactivitiesinbothcelllines.Inspiteofthesesimilarities.J774cellspossessedbothbasicandacidicformsofPKCactivities,whileP388D1cellsownedonlyPKCofbasicform,Nevertheless,thequestionwhyJ774cellsbutnotP388D1cells,canacquirethetumoricidalactivity,aganistP815,cellsfollowingLPStreatmentrematinstobeanswered.
简介:Amurinemacrophage-likecellline,J774,acquried,inresponsetoLPS,anabilitytokilltumornecrosisfactor(TNF)-insensitivetargetP815mastocytomacells,whereasanothercellline,P388D1,didnot.LPS-triggeredsignalingmechanismsbetweenthetwocelllineswerecomparedwithanaimtoinquireaboutthepossiblenatureoftheabove-mentioneddifference.TheresultsshowedthattwocelllinesrespondtoLPS-treatmentbyparallelactivationofbothphospholipasesCandA2(PLCandPLA2)toapproximatelythesameextent.ThemaximumresponseofbothenzymesofJ774cellswasnotedwithin10minofthetreatment,whereasthatofP388D1cellsrequiredmorethan20min.TheotherpropertiesofLPS-responsiveenzymesstudiedweresimilarbetweentwocelllines,ineludingActivationofPLCandPLA2andPKCinmacrophagesbyLPSCa2+augmentationofenzymeactivation,participationofguaninenucleotidebinding(G)proteinsintheinitialactivationprocesses,andinhibitionofenzymeactivationbythepriortreatmentofcellswithcholeraorpartussistoxinsetc.Moreover,LPS-triggeredactivationofPLCandPLA2wasfoundtobefollowedbytheincreaseofPKCactivitiesinbothcelllines.Inspiteofthesesimilarities,J774cellspossessedbothbasicandacidicformsofPKCactivities,whileP388D1cellsownedonlyPKCofbasicform.Nevertheless,thequestionwhyJ774cells,butnotP388D1cells,canacquirethetumoricidalactiyity,aganistP815cellsfollowingLPS-treatmentremainstobeanswered.
简介:TheundecapeptidesubstanceP(SP)wasshowntobeintimatelyinvolvedinboththestructuralandfunctionalaspectsoftheanteriorpituitary.Yet,inadditiontoitsinfluencesonhormonalsecretion,SPmaywellpossessmoreactionsinthismastergland.ThepresentstudywasfthereforeaimedtoinvestigatetheeffectofSPontheproliferationofratanteriorpituitarycellsinprimaryculture,ItwasfoundthatSPcoulddose-dependentlyincreasetheincorporationoftritiatedthymidine(3H-TdR)intoculturedanteriorpituitarycells.OthermammaliantachykininssuchasneurokininAandneurokininBhadsimilareffectbuttovaryingdegrees.TheequipotentanalogueofSP,Norleucine^11-SP(Nle^11-SP),alsoactedlikewise.withitsactionantagonizablebyspantide,aSPreceptorblocker.TofurthercharacterizethenatureofcellsresponsivetothechallengeofSP,immunocytochemicalstainingagainstS-100proteinandsomeadenohypophysealhormoneswasperformedaloneorplusautoradiography.TheresultsshowedthatthepercentageofS-100proteinimmunorectivecellswasapparentlyelevatedbytheaddtionofNle^11-Spfor48h,whichindicatesapreferentialproliferationoffolliculo-stellatecellsundertheregime.ThiswasconfirmedbyincreasesinimmunocytochemicalorautoradiographicallabellingindicesofanteriorpituitarySubstancePandanteriorpituitarycellproliferation.Cellstreatedsimilarly.Takentogether,TheseresultsrevealthatthetrophicactionofSPobservedpreviouslyinothertissuesisalsopresentatleastinculturedratanteriorpituitarycells.withrespondingcellsbeingpredominantlyfolliculo-stellatecellsastypifiedbyS-100proteinimmunoreactivity.Therefore,anintra-pituitarytrophicactionofSPinvivocouldbeanticipated.
简介:cAMPmediatedsignalingmayplayasuppressiveroleinimmuneresponse.WepreviouslyfoundthatthecAMP-elevators(CTxand8-Br-cAMP)inhibitedIL-12,IL-la,IL-6geneexpression,butincreasedthetranscriptionallevelsofIL-10andIL-1RainLPS-treatedmurineperitonealmacrophages.ThepresentstudyexaminedapossiblemolecularmechanisminvolvedincAMPelevators-inducedinhibitionofIL-12p40expressioninresponsetoLPS.OurdatademonstratedthatcAMPelevatorsdownregulatedIL-12p40mRNAexpressionandIL-12pT0productioninmurineperitonealmacrophages.SubsequentstudiesrevealedthatcAMP-elevatorsblockedphosphorylationofp38MAPK,butdidnotaffecttheactivityofNF-κBbindingtoIL-12promoter(-136/-112).ThisisthefirstreportthatcAMPelevatorsinhibitLPS-inducedIL-12productionbyamechanismthatisassociated,atleastinpart,withp38-dependentinhibitionbycAMPsignalingpathways.
简介:TherearetwopossibleoutcomeswhenDNAdamageoccursinnormalmammaliancells:eitherinductionofcell-cyclecheckpointwhichinhibitstheprogressofthecellcyclesaswellasactivatesDNArepairpathways,oractivationofapoptosistoeliminatedamagedcells.Thep53tumour-suppressorgeneplaysakeyroleinselectingthesepathways.Inourpresentworks,thehumangastriccancercelllineAGSwastreatedwithtripchlorolide,apotentantitumorcompoundpurifiedfromaChineseherbTripterygiumWilfordiiHook.Singlecellgelelectrophoresis(Cometassay)showedthatthetreatmentoftripchlorolideresultedinDNAdamageinAGScells.ThedamagedAGScellswentthroughapoptosis,whichwastime-anddose-dependent.
简介:Recentstudiesindicatethatcell-cyclecheckpointsaretightlycorrelatedwiththeregulationofapoptosis,inwhichp53playsanimportantrole.OurpresentworksshowthattheexpressionofE6/E7oncogenesofhumanpapillomavirusinHeLacellsisinhibitedinthepresenceofanti-tumorreagenttripchlorolide(TC),whichresultsintheup-regulationofp53inHeLacells.Interestingly,underthesameTC-treatment,thecellsattheearlyS-phasearemoresusceptibletoapoptosisthanthoseatthemiddleS-phasealthoughp53proteinisstabilizedtothesamelevelinbothsituations.Significantdifferenceisexhibitedbetweenthetwospecifiedexpressionprofiles.Furtheranalysisdemonstratesthatanti-apoptoticgenesurvivinisup-regulatedbyp53intheTC-treatedmiddle-Scells,whereasitisdown-regulatedbyp53intheTC-treatedearly-Scells.Takentogether,thepresentstudyindicatesthatthedifferentialp53-regulatedexpressionofsurvivinatdifferentstagesofthecellcycleresultsindifferentcellularoutputsunderthesameapoptosis-inducer.
简介:CREB有约束力的蛋白质(CBP)和它的相当或相同事物p300是涉及细胞的活动的一个宽数组的各种各样的顺序特定的抄写因素的transcriptionalco使活跃之物,例如脱氧核糖核酸修理,房间生长,区别和apoptosis。Severalstudies建议了CBP和p300可能被看作瘤压制ors,与他们是的突出的角色响应各种各样的刺激的不同基因表示模式的跨coupling。他们主要经由乙酰化施加行动嘘和另外的规章的蛋白质(例如p53)。在CBP/p300功能的主要悖论是他们似乎能够贡献各种各样的反对细胞的进程。呼吸上皮tumorigenesis代表一个范围的多步累积的一个复杂过程基因并且epigenetic错误。通过激活和压抑的建筑群的交替的形成的Transcriptionmodulation是这些精神错乱的最终的收敛点,并且CBP/p300在这相互影响代表关键参加者。因此,他们的分子的行动和相互作用的照明能在呼吸上皮carcinogenesis为药理学干预揭示新潜在的目标。
简介:Brassinosteroids(BR)被transmembrane受体和戏察觉在植物生长和开发的重要角色,以及响应环境刺激的房间。transmembrane受体BRI1能直接绑在brassinolide(BL),并且BAK1与BRI1交往提高发信号的调停BRI1的BR。我们的以前的研究显示了那膜类固醇绑定蛋白质(MSBP1)1能在vitro绑在BL并且否定地涉及BR发信号。进一步阐明内在的机制,我们这里证明MSBP1明确地以一种BL独立的方式在vivo与BAK1的细胞外的领域交往。由MSBP1的增加的表示的压制的房间扩大和BR回答能被overexpressingBAK1或它的细胞内部的kinase领域恢复,建议MSBP1可以压制通过与BAK1交往发信号的BR。Subcellular本地化研究表明MSBP1和BAK1对血浆膜和endocytic泡局部性,MSBP1加速BAK1endocytosis,它导致由向内涵体转移BAK1的平衡发信号的压制的BR。确实,提高了MSBP1表示还原剂在在vivo的BRI1和BAK1之间的相互作用,表明那MSBP1在发信号的BR的早步充当一个否定因素小径。
简介:Thec-erbB-2proto-oncogeneencodesa185kDaproteinp185,whichbelongstoepidermalgrowthfactorreceptorfamily.Amplificationofthisgenehasbeenshowntocorrelatewithpoorclinicalprognosisforcertaincancerpatients.ThemonoclonalantibodyA21whichdirectedagainstp185specificallyinhibitsproliferationoftumorcellsoverexpressingp185,henceallowsittobeacandidatefortargetedtherapy.InordertoovercomeseveraldrawbacksofmurineMAb,wecloneditsVHandVLgenesandconstructedthesingle-chainFv(scFv)throughapeptidelinker.TherecombinantscFvA21wasexpressedinEscherichiacoliandpurifiedbytheaffinitycolumn.SubsequentlyitwascharacterizedbyELISA,Westernblot,cellimmunohistochemistryandFACS.Alltheseassaysshowedthebindingactivitytoextracellulardomain(ECD)ofp185.BasedonthosepropertiesofscFvA21,wefurtherconstructedthescFv-Fcfusionmoleculewithahomodimerformandtherecombinantproductwasexpressedinmammaliancells.Inaseriesofsubsequentanalysisthisfusionproteinshowedidenticalantigenbindingsiteandactivitywiththeparentantibody.Theseanti-p185engineeredantibodieshavepromisedtobefurthermodifiedasatumortargetingdrugs,withaviewofapplicationinthediagnosisandtreatmentofhumanbreastcancer.
简介:Arabidopsisthaliana嘘一deacetylase1(AtHD1或AtHDA19),酵母RPD3的一个相当或相同的事物,是在植物的许多生理、发展的过程的一个全球管理者。尽管有为在植物基因规定和开发的AtHD1的一个角色的基因证据,AtHD1的生物化学、细胞的性质糟糕被理解。这里,我们在vivo报导AtHD1的细胞的本地化模式并且嘘在vitro的一项deacetylase活动。绿荧光灯的蛋白质(GFP)的短暂、稳定的表示在洋葱房间标注了AtHD1并且分别地,在转基因的Arabidopsis的根,种子和叶子表明AtHD1在euchromatic区域大概在原子核是局部性的并且从核排除。AtHD1的本地化模式与涉及核形成和transgenes的silencing并且分别地重复了DNA元素的AtHD2和AtHDA6的那些不同。另外,一hist一deacetylase活动试金证明在细菌生产的recombinantAtHD1示威了一特定嘘在vitro的一项deacetylase活动。数据建议AtHD1是原子蛋白质并且拥有嘘为对植物生长和开发重要的全球transcriptional规定负责的一项deacetylase活动。
简介:它一致地被看了那oncoproteinp28GANK,它是在人的hepatocellular癌(HCC)的overexpressed,在HCC的tumorigenesis起一个关键作用。然而,内在的机制仍然保持不清楚。这里,我们证明p28GANK在endoplasmic蜂窝胃导致的HCC房间禁止apoptosis(嗯)应力。在期间嗯应力,p28GANK提高展开的蛋白质反应,支持嗯从翻译压抑的恢复,并且从而便于房间的能力应付压力条件。而且,p28GANKupregulates调整葡萄糖的蛋白质78(GRP78),一把钥匙嗯女伴蛋白质,它随后提高合拢能力的ER并且支持恢复从嗯应力。我们也证明p28GANK增加p38激活mitogen的蛋白质kinase和Aktphosphorylation,并且禁止原子因素kappaB(NF-B)激活在下面嗯强调upregulation,它接着贡献GRP78。一起拿,我们的结果显示p28GANK禁止嗯在HCC房间的导致压力的apoptosis,至少部分地,由提高适应反应和GRP78表示。我们建议p28GANK在ER压力条件下面为HCC前进有潜在的含意。