Gang Fu / Lei Zhao / Erin Dymek / Yuqing Hou / Kangkang Song / Nhan Phan / Zhiguo Shang / Elizabeth F Smith / George B Witman / Daniela Nicastro /
PubMed Abstract
Nearly all motile cilia contain a central apparatus (CA) composed of two connected singlet microtubules with attached projections that play crucial roles in regulating ciliary motility. Defects in CA ...Nearly all motile cilia contain a central apparatus (CA) composed of two connected singlet microtubules with attached projections that play crucial roles in regulating ciliary motility. Defects in CA assembly usually result in motility-impaired or paralyzed cilia, which in humans causes disease. Despite their importance, the protein composition and functions of the CA projections are largely unknown. Here, we integrated biochemical and genetic approaches with cryo-electron tomography to compare the CA of wild-type with CA mutants. We identified a large (>2 MD) complex, the C1a-e-c supercomplex, that requires the PF16 protein for assembly and contains the CA components FAP76, FAP81, FAP92, and FAP216. We localized these subunits within the supercomplex using nanogold labeling and show that loss of any one of them results in impaired ciliary motility. These data provide insight into the subunit organization and 3D structure of the CA, which is a prerequisite for understanding the molecular mechanisms by which the CA regulates ciliary beating.
EMDB-20160: The central pair apparatus focusing on the C1a-e-c supercomplex extracted from the cryo-electron tomography and subtomographic average of isolated Chlamydomonas wild type axoneme Method: EM (subtomogram averaging) / Resolution: 23 Å
EMDB-20161: The central pair apparatus focusing on the C1a-e-c supercomplex extracted from the cryo-electron tomography and subtomographic average of isolated Chlamydomonas fap92 mutant axoneme Method: EM (subtomogram averaging) / Resolution: 22 Å
EMDB-20162: The central pair apparatus focusing on the C1a-e-c supercomplex extracted from the cryo-electron tomography and subtomographic average of isolated Chlamydomonas fap76-1 mutant axoneme Method: EM (subtomogram averaging) / Resolution: 22 Å
EMDB-20163: The central pair apparatus focusing on the C1a-e-c supercomplex extracted from the cryo-electron tomography and subtomographic average of isolated Chlamydomonas fap76-2 mutant axoneme Method: EM (subtomogram averaging) / Resolution: 26 Å
EMDB-20164: The central pair apparatus focusing on the C1a-e-c supercomplex extracted from the cryo-electron tomography and subtomographic average of isolated Chlamydomonas fap81 mutant axoneme Method: EM (subtomogram averaging) / Resolution: 22 Å
EMDB-20165: The central pair apparatus focusing on the C1a-e-c supercomplex extracted from the cryo-electron tomography and subtomographic average of isolated Chlamydomonas fap216 mutant axoneme Method: EM (subtomogram averaging) / Resolution: 22 Å
EMDB-20166: The central pair apparatus focusing on the C1a-e-c supercomplex extracted from the cryo-electron tomography and subtomographic average of isolated Chlamydomonas fap76-1,fap81 double mutant axoneme Method: EM (subtomogram averaging) / Resolution: 23 Å
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