Genome-Wide Analysis of Binding Sites and Direct Target Genes of the Orphan Nuclear Receptor NR2F1COUP-TFI 英文参考文献.docVIP
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Genome-Wide Analysis of Binding Sites and Direct Target Genes of the Orphan Nuclear Receptor NR2F1COUP-TFI 英文参考文献
Genome-WideAnalysisofBindingSitesandDirect
TargetGenesoftheOrphanNuclearReceptorNR2F1/
COUP-TFI
CelinaMontemayor1,2,OscarA.Montemayor2,AlexRidgeway1,FengLin2,DavidA.Wheeler4,5,6,ScottD.
Pletcher2,4,5,FredA.Pereira1,2,3,4,7
*
1DepartmentofMolecularandCellularBiology,BaylorCollegeofMedicine,Houston,Texas,UnitedStatesofAmerica,2HufftingtonCenteronAging,BaylorCollegeof
Medicine,Houston,Texas,UnitedStatesofAmerica,3BobbyR.AlfordDepartmentofOtolaringology–HeadandNeckSurgery,BaylorCollegeofMedicine,Houston,
Texas,UnitedStatesofAmerica,4PrograminCellandMolecularBiology,BaylorCollegeofMedicine,Houston,Texas,UnitedStatesofAmerica,5DepartmentofHuman
andMolecularGenetics,BaylorCollegeofMedicine,Houston,Texas,UnitedStatesofAmerica, 6HumanGenomeCenter,BaylorCollegeofMedicine,Houston,Texas,
UnitedStatesofAmerica,7DepartmentofBioengineering,RiceUniversity,Houston,Texas,UnitedStatesofAmerica
Abstract
Background: Identification of bona fide direct nuclear receptor gene targets has been challenging but essential for
understandingregulationoforganismalphysiologicalprocesses.
Results:Wedescribeamethodologytoidentifytranscriptionfactorbindingsitesandtargetgenesinvivobyintersecting
microarray data, computational binding site queries, and evolutionary conservation. We provide detailed experimental
validationofeachstepand,asaproofofprinciple,utilizethemethodologytoidentifynoveldirecttargetsoftheorphan
nuclearreceptorNR2F1(COUP-TFI).Thefirststepinvolvedvalidationofmicroarraygeneexpressionprofilesobtainedfrom
wild-type and COUP-TFI2/2 inner ear tissues. Secondly, we developed a bioinformatic tool to search for COUP-TFI DNA
bindingsitesingenomes,usingaclassification-typeHiddenMarkovModeltrainedwith49publishedCOUP-TFresponse
elements.Wenextobtainedarankedlistofcandidate invivodirectCOUP-TFItargetsbyintegratingthemicroarrayand
bioinformatics analyses according to the degree of binding site evolutionary conservation and microarray statistical
significance.Lastly,asproof-of-concept,5s
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