Dysferlin Interacts with Tubulin and Microtubules in Mouse Skeletal Muscle 英文参考文献.docVIP

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Dysferlin Interacts with Tubulin and Microtubules in Mouse Skeletal Muscle 英文参考文献.doc

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Dysferlin Interacts with Tubulin and Microtubules in Mouse Skeletal Muscle 英文参考文献

DysferlinInteractswithTubulinandMicrotubulesin MouseSkeletalMuscle BilalA.Azakir1,SabrinaDiFulvio1,ChristianTherrien1,MichaelSinnreich1,2* 1NeuromuscularResearchGroup,MontrealNeurologicalInstituteandHospital,McGillUniversity,Montreal,Quebec,Canada,2NeuromuscularCenter,Departmentsof NeurologyandBiomedicine,UniversityHospitalBasel,Basel,Switzerland Abstract DysferlinisatypeIItransmembraneproteinimplicatedinsurfacemembranerepairinmuscle.Mutationsindysferlinleadto limbgirdlemusculardystrophy2B,MiyoshiMyopathyanddistalanteriorcompartmentmyopathy.Dysferlin’smodeofactionis notwellunderstoodandonlyafewproteinbindingpartnershavethusfarbeenidentified.Usingaffinitypurificationfollowed by liquid chromatography/mass spectrometry, we identified alpha-tubulin as a novel binding partner for dysferlin. The associationbetweendysferlinandalpha-tubulin,aswellasbetweendysferlinandmicrotubules,wasconfirmedinvitroby glutathione S-transferase pulldown and microtubule binding assays. These interactions were confirmed in vivo by co- immunoprecipitation.Confocalmicroscopyrevealedthatdysferlinandalpha-tubulinco-localizedintheperinuclearregion andinvesicularstructuresinmyoblasts,andalongthinlongitudinalstructuresreminiscentofmicrotubulesinmyotubes.We mappeddysferlin’salpha-tubulin-bindingregiontoitsC2AandC2Bdomains.Modulationofcalciumlevelsdidnotaffect dysferlin binding to alpha-tubulin, suggesting that this interaction is calcium-independent. Our studies identified a new bindingpartnerfordysferlinandsuggestaroleformicrotubulesindysferlintraffickingtothesarcolemma. Citation:AzakirBA,DiFulvioS,TherrienC,SinnreichM(2010)DysferlinInteractswithTubulinandMicrotubulesinMouseSkeletalMuscle.PLoSONE5(4):e10122. doi:10.1371/journal.pone.0010122 Editor:NeilA.Hotchin,UniversityofBirmingham,UnitedKingdom ReceivedDecember1,2009;AcceptedMarch11,2010;PublishedApril12,2010 Copyright: ? 2010 Azakir et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, wh