Broad Expression Analysis of Human ANTXR1TEM8 Transcripts Reveals Differential Expression and Novel Splizce Variants 英文参考文献.docVIP

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Broad Expression Analysis of Human ANTXR1TEM8 Transcripts Reveals Differential Expression and Novel Splizce Variants 英文参考文献.doc

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Broad Expression Analysis of Human ANTXR1TEM8 Transcripts Reveals Differential Expression and Novel Splizce Variants 英文参考文献

BroadExpressionAnalysisofHumanANTXR1/TEM8 TranscriptsRevealsDifferentialExpressionandNovel SplizceVariants MicaelaVargas1¤,RaghavendraKaramsetty1,StephenH.Leppla2,G.JilaniChaudry1* 1Cell and Molecular Biology Program, Department of Biology, The University of Texas at San Antonio, San Antonio, Texas, United States of America, 2Microbial PathogenesisSection,TheLaboratoryofParasiticDiseases,NationalInstituteofAllergyandInfectiousDiseases,NationalInstitutesofHealth,Bethesda,Maryland,United StatesofAmerica Abstract Tumorendothelialmarker8(TEM8;ANTXR1)isoneoftwoanthraxtoxinreceptors;theotheriscapillarymorphogenesis gene 2 protein (CMG2; ANTXR2). TEM8 shows enhanced expression in certain tumor endothelia, and is thought to be aplayerintumorvasculatureformation.However,acomprehensiveexpressionprofileofindividualTEM8variantsinnormal orcanceroustissuesislacking.InthisworkwecarriedoutanextensiveanalysisofallsplicevariantsofhumanTEM8in12 digestive tissues, and 8 each fetal and adult tissues, 6 of them cognate pairs. Using variant-specific primers, we first ascertainedthestatusoffull-lengthtranscriptsbynestedPCR.Wethencarriedoutquantitativeanalysisofeachtranscript byreal-timePCR.ThreesplicevariantsofTEM8werereportedbefore,twosingle-passintegralmembraneforms(V1andV2) andonesecreted(V3).Ouranalysishasrevealedtwonewvariants,oneencodingamembrane-boundformofthereceptor and the other secreted, which we have designated V4 and V5, respectively. All tissues had V1, V2, V3, and V4, but only prostatehadV5.Real-timePCRrevealedthatallvariantsarepresentatdifferentlevelsinvarioustissues.V3appearedthe mostabundantofall.Toascertainitsfunctionalityforanthraxtoxin,weexpressedthenewlyidentifiedformV4inareceptor- negativehostcell,andincludedV1andV2forcomparison.Cytotoxicity,toxinbinding,andinternalizationassaysshowedV4 tobeasefficientareceptorasV1andV2. Citation:VargasM,KaramsettyR,LepplaSH,ChaudryGJ(2012)BroadExpressionAnalysisofHumanANTXR1/TEM8TranscriptsRevealsDifferentialExpression andNovelSpl