Protein-Binding Microarray Analysis of Tumor Suppressor AP2α Target Gene Specificity 英文参考文献.docVIP

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Protein-Binding Microarray Analysis of Tumor Suppressor AP2α Target Gene Specificity 英文参考文献.doc

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Protein-Binding Microarray Analysis of Tumor Suppressor AP2α Target Gene Specificity 英文参考文献

Protein-BindingMicroarrayAnalysisofTumor SuppressorAP2aTargetGeneSpecificity JanKerschgens1.,Ste′phanieRenaud1.,Fre′de′ricSchu¨tz2,LuiginoGrasso3,TanjaEgener-Kuhn1,4 ,Jean- Franc?oisDelaloye5,Hans-AntonLehr5,HorstVogel3,NicolasMermod1* 1Institute of Biotechnology, University of Lausanne, Lausanne, Switzerland, 2Swiss Institute of Bioinformatics, University of Lausanne, Lausanne, Switzerland, 3LaboratoryofPhysicalChemistryofPolymersandMembranes,E′colePolytechniqueFe′de′raledeLausanne,Lausanne,Switzerland,4NationalCompetenceCenterfor BiomedicalImaging,E′colePolytechniqueFe′de′raledeLausanne,Lausanne,Switzerland,5CentreHospitalierUniversitaireVaudois,Lausanne,Switzerland Abstract Cheap and massively parallel methods to assess the DNA-binding specificity of transcription factors are actively sought, given their prominent regulatory role in cellular processes and diseases. Here we evaluated the use of protein-binding microarrays (PBM) to probe the association of the tumor suppressor AP2a with 6000 human genomic DNA regulatory sequences. We show that the PBM provides accurate relative binding affinities when compared to quantitative surface plasmonresonanceassays.APBM-basedstudyofhumanhealthyandbreasttumortissueextractsallowedtheidentification ofpreviouslyunknownAP2atargetgenesanditrevealedgeneswhosedirectorindirectinteractionswithAP2aareaffected in the diseased tissues. AP2a binding and regulation was confirmed experimentally in human carcinoma cells for novel targetgenesinvolvedintumorprogressionandresistancetochemotherapeutics,providingamolecularinterpretationof AP2aroleincancerchemoresistance.Overall,weconcludethatthisapproachprovidesquantitativeandaccurateassaysof the specificity and activity of tumor suppressor and oncogenic proteins in clinical samples, interfacing genomic and proteomicassays. Citation:KerschgensJ,RenaudS,Schu¨tzF,GrassoL,Egener-KuhnT,etal.(2011)Protein-BindingMicroarrayAnalysisofTumorSuppressorAP2aTargetGene Specificity.PLoSONE6(8):e22895.doi:10