One of the Nine Doublet Microtubules of Eukaryotic Flagella Exhibits Unique and Partially Conserved Structures 英文参考文献.docVIP

OneoftheNineDoubletMicrotubulesofEukaryotic FlagellaExhibitsUniqueandPartiallyConserved Structures JianfengLin,ThomasHeuser,KangkangSong,XiaofengFu,DanielaNicastro* BiologyDepartment,RosenstielCenter,BrandeisUniversity,Waltham,Massachusetts,UnitedStatesofAmerica Abstract The axonemal core of motile cilia and flagella consists of nine doublet microtubules surrounding two central single microtubules. Attached to the doublets are thousands of dynein motors that produce sliding between neighboring doublets,whichinturncausesflagellarbending.Althoughmanystructuralfeaturesoftheaxonemehavebeendescribed, structuresthatareuniquetospecificdoubletsremainlargelyuncharacterized.Thesedoublet-specificstructuresintroduce asymmetry intotheaxoneme andarelikelyimportantfor thespatial control oflocalmicrotubulesliding. Here,weused cryo-electron tomography and doublet-specific averaging to determine the 3D structures of individual doublets in the flagella of two evolutionarily distant organisms, the protist Chlamydomonas and the sea urchin Strongylocentrotus . We demonstratethat,inbothorganisms,oneoftheninedoubletsexhibitsuniquestructuralfeatures.Someofthesefeatures arehighlyconserved,suchastheinter-doubletlinki-SUB5-6,whichconnectsthisdoublettoitsneighborwithaperiodicity of 96nm. We also show that the previously described inter-doublet links attached to this doublet, the o-SUB5-6 in Strongylocentrotus andtheproximal 1–2bridge in Chlamydomonas,arelikelynot homologousfeatures. The presence of inter-doublet links and reduction of dynein arms indicate that inter-doublet sliding of this unique doublet against its neighborislimited,providingarigidplaneperpendiculartotheflagellarbendingplane.Thesedoublet-specificfeaturesand the non-sliding nature of these connected doublets suggest a structural basis for the asymmetric distribution of dynein activityandinter-doubletsliding,resultinginquasi-planarwaveformstypicalof9+2ciliaandflagella. Citation:LinJ,HeuserT,SongK,FuX,NicastroD(2012)Oneoft