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TitleStructural organization of the C1a-e-c supercomplex within the ciliary central apparatus.
Journal, issue, pagesJ Cell Biol, Vol. 218, Issue 12, Page 4236-4251, Year 2019
Publish dateDec 2, 2019
AuthorsGang Fu / Lei Zhao / Erin Dymek / Yuqing Hou / Kangkang Song / Nhan Phan / Zhiguo Shang / Elizabeth F Smith / George B Witman / Daniela Nicastro /
PubMed AbstractNearly 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.
External linksJ Cell Biol / PubMed:31672705 / PubMed Central
MethodsEM (subtomogram averaging)
Resolution22 - 26 Å
Structure data

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

Source
  • Chlamydomonas reinhardtii (plant)
KeywordsAxoneme / Cell Movement / Chlamydomonas / Cilia / Electron Microscope Tomography / Flagella / Genotype / Microtubules / Molecular Conformation / Mutation / Phenotype / Polymerase Chain Reaction

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