简介：Wntsignalingplaysanimportantroleinembryogenesisandtumorgenesis.AlthoughthemechanismabouthowWntstransducetheirsignalingfromreceptorfrizzled(Fz)tocytosolhasnotbeenunderstood,dishevelled(Dvl)proteinwasconsideredastheintersectionofWntsignaltraffic.Inthisstudy,wecharacterizedthefunctionofthreedomains(DIX,PDZandDEP)ofDvl-1incanonicalWntsignaltransductionandDvl-1membranetranslocation.ItwasfoundbothDIXandDEPdomainweresufficienttoblockWnt-3a-inducedLEF-1transcriptionalactivityandfreecytosolβ-cateninaccumulation;whereasPDZdomainandafunctionalmutantformofDEPdomain(DEP-KM)hadnoeffectoncanonicalWntsignaling.Inaddition,whencotransfectedwithFz-7,DEPdomain,butnotDIX,PDZorDEP-KM,translocatedandco-localizedwithFz-7totheplasmamembrane,whichwassimilartoDvl-1.Furthermore,itwasDEPdomainthatcouldblockFz-7-inducedmembranetranslocationofDvl-1viaapossiblecompetitivemechanism.TheseresultsstronglysuggestthatDEPdomainisresponsibleforthemembranetranslocationofDvl-1proteinuponWntsignalstimulation.

简介：Glucosetransporter4(GLUT4)isresponsibleforinsulin-stimulatedglucosetransportingintotheinsulin-sensitivefatandmusclecells.ThedynamicsofGLUT4storagevesicles(GSVs)remainstobeexploredanditisunclearhowGSVsarearrangedbasedontheirmobility.Weexaminedthisissuein3T3-L1cellsviainvestigatingthethree-dimensionalmobilityofsingleGSVlabeledwithEGFP-fusedGLUT4.Athinlayerofcytosolrightadjacenttotheplasmamembranewasilluminatedandsuccessivelyimagedat5Hzunderatotalinternalreflectionfluorescencemicroscopewithapenetrationdepthof136nm.Employingsingleparticletracking,thethree-dimensionalsubpixeldisplacementofsingleGSVwastrackedataspatialprecisionof22nm.Boththemeansquaredisplacementandthediffusioncoefficientwerecalculatedforeachvesicle.Trackingresultsrevealedthatvesiclesmovedasifrestrictedwithinacagethathasameanradiusof160nm,suggestingthepresenceofsomeintracellulartetheringmatrix.ByconstructingthehistogramofthediffusioncoefficientsofGSVs,weobservedasmoothdistributioninsteadoftheexistenceofdistinctgroups.TheresultindicatesthatGSVsaredynamicallyretainedinacontinuousandwiderangeofmobilityratherthanintoseparateclasses.

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：<正>Uponactivation,naiveT-helpercellscandifferentiateintotwomajordistinctsubsets,Thelper1(Th1)andThelper2(Th2),asdefinedbytheireffectorfunctionsandcytokinesecretionpatterns.CytokinemilieuandcostimulatorymoleculeshavebeenshowntoplayanessentialroleindeterminingThelperdifferentiation.However,itisstillunclearhowtheeffectsofsignalsofco-stimulatorymoleculesandcytokinesareexertedduringThelperdifferentiation.Weshowevidencesuggestingthatwhilecytokinesignalsinitiatedifferentiationprogram,theselectiveactionofdeatheffectorsdeterminestheendpointbalanceofdifferenti-

简介：AktandBcl-xLbothpromoteresistancetoapoptosis.AcomparisonofAkt-andBcl-xL-dependentcellsurvivalwasundertaken.ExpressionofconstitutivelyactiveAktallowscellstosurviveforprolongedperiodsintheabsenceofgrowthfactors.ThissurvivalcorrelateswiththeexpressionlevelofactivatedAktandiscomparableinmagnitudetotheprotectionprovidedbytheanti-apoptoticgeneBcl-xL.Althoughbothgenespreventcelldeath,Akt-protectedcellscanbedistinguishedfromBcl-xL-protectedcellsonthebasisofincreasedglucosetransporterexpression,glycolyticactivity,mitochondrialpotential,andcellsize.Inaddition,Akt-expressingcellsrequirehighlevelsofextracellularnutrientstosupportcellsurvival.In

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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-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：ThemuscleproteinmyosinbindingproteinC(MyBPC)isalargemulti-domainproteinwhoseroleinthesarcomereiscomplexandnotyetfullyunderstood.MutationsinMyBPCarestronglyassociatedwiththeheartdiseasefamilialhypertrophiccardiomyopathy(FHC)andtheseexperimentsofnaturehaveprovidedsomeinsightintotheintricateworkingsofthisproteinintheheart.WhilesomeregionsoftheMyBPCmoleculehavebeenassignedafunctionintheregulationofmusclecontraction,theinteractionofotherregionswithvariouspartsofthemyosinmoleculeandthesarcomericproteins,actinandtitin,remainobscure.Inadditicn,severalintra-domaininteractionsbetweenadjacentMyBPCmoleculeshavebeenidentified.Althoughthebasicstructureofthemolecule(aseriesofimmunoglobulinandfibronectindomains)hasbeenelucidated,theassemblyofMyBPCinthesarcomereisatopicfordebate.ByanalysingtheMyBPCsequencewithrespecttoFHC-causingmutationsitispossibletoidentifyindividualresiduesorregionsofeachdomainthatmaybeimportanteitherforbindingorregulation.Thisreviewlooksatthecurrentliterature,inconcertwithalignmentsandthestructuralmodelsofMyBPC,inanattempttounderstandhowFHCmutationsmayleadtothediseasestate.

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：Changesinthedistributionof1P1-antigeninthedevelopingchickretinahavebeenexaminedbyindriectimmunofluorescencestainingtechniqueusingthenovelmonoclonalantibody(MAb)1P1.Expressionofthe1P1antigenwasfoundtoberegulatedinradialaswellasintangentialdimensionoftheretina,beingpreferentiallyorexclusivelylocatedintheinnerandouterplexiformlayersoftheneuralretinadependingonthestagesofdevelopment,Withtheonsetoftheformationoftheinnerplexiformlayer1P1antigenbecomesexpressedintheretina.Withprogressingdifferentiationoftheinnerplexiformlayer1P1immunofluorescencerevealed2subbandsatE9and6subandsatE18,Atpostnatalstages(afterP3)immunoreactivitywasreducedinaninside-outsidesequenceleadingtothecompleteabsenceofthe1P1antigeninadulthood.1P1antigenexpressionintheouterplexiformlayerwasalsosubjecttodevelopmentalregulation.Thespation-temporalpatternof1P1antigenexpressionwascorrelatedwiththetimecourseofhistologicaldifferentationofchickretina,namelythesynapserichplexiformlayers.Whetherthe1P1antigenwasfunctionallyinvolvedindendriteextensionandsynapseformationwasdiscussed.

简介：铜绑定，抛锚膜的、细胞的prion蛋白质(PrPC)有二个组成的劈开地点生产不同N终端和C终端碎片(N1/C1和N2/C2)。用表示也人的PrPC，鼠标PrPC或鼠标PrPC的RK13房间带3F4epitope，这研究在endoproteolytic劈开和一个通常认为的PrPC函数上探索了PrPC主要顺序的影响，地图kinase信号transduction，响应外长的铜与或没有使不安的膜环境。PrPC主要顺序，特别在N1/C1劈开地点附近，看起来在这个地点和细胞外的调整信号的kinase1/2(ERK1/2)phosphorylation影响解朊作用的基础层次，与处理激活增加与基础ERK1/2表明一种反的关系。人的PrPC独自响应铜显示出增加的N1/C1劈开，由特定的p38和JNK/SAPKphosphorylation伴随了。到加扣押胆固醇的抗菌素filipin的铜的联合暴露导致了一只老鼠信号蛋白质phosphorylation的PrPC特定的实质的增加，由N1/C1劈开的增加伴随了。怀有人的N1/C1劈开地点的老鼠PrPC基础地并且响应铜假定更似人类的侧面并且改变了膜环境。我们的结果证明在N1/C1劈开地点附近的PrPC主要顺序影响在这个地点处理的endoproteolytic，它显得连接了基础地并且响应铜印射kinase信号transduction。进一步，主要顺序看起来响应外长的刺激在PrPC相关的信号transduction的忠实上授与N1/C1劈开和膜完整的相互的依赖。

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：Thenorepinephrinetransporter(NET)isamemberoftheNa^+/Cl^-dependentneurotransmittertransporterfamilyandconstitutesthetargetofseveralclinicallyimportantantidepressants.TodelineatethecriticalaminoacidresiduesandthefunctionofC-terminalinregulatingtransportactivityofNET,hereweconstructedtwositemutants(V70F,F72V;V70I,F72V)andoneC-terminaltruncatedmutant(Δ611-617).ThewildtypeandmutantsofNETwereexpressedinXenopusoocytesbyinjectionoftheircRNA.Wefoundthatallofthesemutantslosttheirtransportactivity.TheseresultsindicatethattheaminoacidresiduesofV70andF72,andthelastsevenaminoacidsofC-terminalareessentialtothetransportactivityofNET.

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.

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简介：TheyeastHAL1genewasintroducedintoArabidopsisthalianabyAgrobacteriumtumefaciens-mediatedtransformationwithvacuuminfiltrationunderthecontrolofCaMV35Spromoter.Thirty-threeindividualkanamycinresistantplantswereobtainedfrom75,000seeds.SouthernblottinganalysisindicatedthatHAL1genehadbeenintegratedintoallofthetransgenicplants'genomes.ThecopynumberofHAL1geneintransgenicplantswasmostly1to3bySouthernanalysis.Phenotypesoftransgenicplantshavenodifferenceswithwildtypeplants.Severalsamplesoftransformantswereself-pollinated,andprogeniesfromtransformedandnon-transformedplants(controls)wereevaluatedforsalttoleranceandgeneexpression.MeasurementofconcentrationsofintracellularK+andNa+showedthattransgeniclineswereabletoretainlessNa+thanthatofthecontrolundersaltstress.ResultsfromdifferenttestsindicatedtheexpressionofHAL1genepromotesahigherlevelofsalttoleranceinvivointhetransgenicArabidopsisplants.

简介：Brassinosteroids(BR)是植物激素的一个主要的组调整植物生长和开发。BRI1，对质膜局部性的蛋白质，当BR受体和它被建议了，工作它的kinase活动在调整BR的植物生长和开发有一个必要角色。这里，我们报导隔离和bri1的新等位基因的分子的描述，bri1-301，哪个表演中等词法显型和减少的回答到在正常生长条件下面的BR。顺序分析从GG识别了二底的改变到，导致到在BRI1kinase领域的989I的989G的变换在。kinase活动的试管内试金证明bri1-301不向BRI1底层TTL和BAK1举办可检测的autophosphorylation活动或磷酸化活动。而且，我们的结果建议甚至与极其损害的kinase活动，bri1-301仍然在调整植物生长和开发保留部分功能，它提出BRI1kinase活动是否在高等植物为调停BR的生长和开发是必要的问题。

简介：被象染色体冷凝作用，面向双性人的锭子形成，染色体大会离子，分离和胞质分裂那样的一系列紧安排的事件精确通过有丝分裂调整。染色体运动被和位于着丝点以内的一个专业化染色体领域的锭子微导管的相互作用在有丝分裂期间管理。这个专业化区域，叫了kinetochore，是为锭子微导管着丝点的地点协会。除了用作在染色体和锭子微导管之间的一个物理连接，kinetochore通过微导管马达和定位在或在它附近的锭子集会检查点(囊)传感器在chromosomal分离展出一个活跃角色[例如，1]。