Genome-Scale Identification of Membrane-Associated Human mRNAs 英文参考文献.docVIP

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Genome-Scale Identification of Membrane-Associated Human mRNAs 英文参考文献.doc

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Genome-Scale Identification of Membrane-Associated Human mRNAs 英文参考文献

Genome-ScaleIdentification ofMembrane-AssociatedHumanmRNAs Maximilian Diehn1,2,Ramona Bhattacharya2,David Botstein3,4,Patrick O.Brown2,5* 1 Department of Radiation Oncology, Stanford University School of Medicine, Stanford, California, United States of America, 2 Department of Biochemistry, Stanford UniversitySchoolofMedicine,Stanford,California,UnitedStatesofAmerica,3DepartmentofGenetics,StanfordUniversitySchoolofMedicine,Stanford,California,United States of America, 4 Lewis-Sigler Institute for Integrative Genomics, Princeton University, Princeton, New Jersey, United States of America, 5 Howard Hughes Medical Institute,StanfordUniversitySchoolofMedicine,Stanford,California,UnitedStatesofAmerica Thesubcellularlocalizationofproteinsiscriticaltotheirbiologicalroles.Moreover,whetheraproteinismembrane- bound,secreted,orintracellularaffectstheusefulnessof,andthestrategiesfor,usingaproteinasadiagnosticmarker oratargetfortherapy.Weemployedarapidandefficientexperimentalapproachtoclassifythousandsofhumangene products as either ‘‘membrane-associated/secreted’’ (MS) or ‘‘cytosolic/nuclear’’ (CN). Using subcellular fractionation methods,weseparatedmRNAsassociatedwithmembranesfromthoseassociatedwiththesolublecytosolicfraction andanalyzedthesetwopoolsbycomparativehybridizationtoDNAmicroarrays.Analysisof11differenthumancell lines,representinglymphoid,myeloid,breast,ovarian,hepatic,colon,andprostatetissues,identifiedmorethan5,000 previouslyuncharacterizedMSandmorethan6,400putativeCNgenesathighconfidencelevels.Theexperimentally determinedlocalizationscorrelatedwellwithinsilicopredictionsofsignalpeptidesandtransmembranedomains,but alsosignificantlyincreasedthenumberofhumangenesthatcouldbecatalogedasencodingeitherMSorCNproteins. Usinggeneexpressiondatafromavarietyofprimaryhumanmalignanciesandnormaltissues,werationallyidentified hundredsofMSgeneproductsthataresignificantly overexpressedintumorscompared tonormal tissuesandthus representcandidatesforserumdiagnostictestsormonoclo