Protein Phosphatase-1α Interacts with and Dephosphorylates Polycystin-1 英文参考文献.docVIP

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Protein Phosphatase-1α Interacts with and Dephosphorylates Polycystin-1 英文参考文献.doc

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Protein Phosphatase-1α Interacts with and Dephosphorylates Polycystin-1 英文参考文献

ProteinPhosphatase-1aInteractswithand DephosphorylatesPolycystin-1 StephenC.Parnell1,2*,SanjeevPuri3,DarrenP.Wallace2,4,JamesP.Calvet1,2 * 1DepartmentofBiochemistryandMolecularBiology,UniversityofKansasMedicalCenter,KansasCity,Kansas,UnitedStatesofAmerica,2KidneyInstitute,Universityof KansasMedicalCenter,KansasCity,Kansas,UnitedStatesofAmerica,3BiotechnologyDepartment,UniversityInstituteofEngineeringandTechnology,PanjabUniversity, Chandigarh,India,4DepartmentofMedicineandtheKidneyInstitute,UniversityofKansasMedicalCenter,KansasCity,Kansas,UnitedStatesofAmerica Abstract Polycystin signaling is likely to be regulated by phosphorylation. While a number of potential protein kinases and their target phosphorylation sites on polycystin-1 have been identified, the corresponding phosphatases have not been extensivelystudied.Wehavenowdeterminedthatpolycystin-1isaregulatorysubunitforproteinphosphatase-1a(PP1a). SequenceanalysishasrevealedthepresenceofahighlyconservedPP1-interactionmotifinthecytosolic,C-terminaltailof polycystin-1; and we have shown that transfected PP1a specifically co-immunoprecipitates with a polycystin-1 C-tail construct. To determine whether PP1a dephosphorylates polycystin-1, a PKA-phosphorylated GST-polycystin-1 fusion proteinwasshowntobedephosphorylatedbyPP1abutnotbyPP2B(calcineurin).MutationswithinthePP1-bindingmotif of polycystin-1, including an autosomal dominant polycystic kidney disease (ADPKD)-associated mutation, significantly reduced PP1a-mediated dephosphorylation of polycystin-1. The results suggest that polycystin-1 forms a holoenzyme complex with PP1a via a conserved PP1-binding motif within the polycystin-1 C-tail, and that PKA-phosphorylated polycystin-1servesasasubstratefortheholoenzyme. Citation: Parnell SC, Puri S, Wallace DP, Calvet JP (2012) Protein Phosphatase-1a Interacts with and Dephosphorylates Polycystin-1. PLoS ONE 7(6): e36798. doi:10.1371/journal.pone.0036798 Editor:NickGay,UniversityofCambridge,UnitedKingdom ReceivedApril