TY - JOUR
T1 - Structural organization of the C1a-e-c supercomplex within the ciliary central apparatus
AU - Fu, Gang
AU - Zhao, Lei
AU - Dymek, Erin
AU - Hou, Yuqing
AU - Song, Kangkang
AU - Phan, Nhan
AU - Shang, Zhiguo
AU - Smith, Elizabeth F.
AU - Witman, George B.
AU - Nicastro, Daniela
N1 - Funding Information:
We are grateful to D. Stoddard for management of the UTSW cryo-electron microscope facility, which is funded in part by a Cancer Prevention and Research Institute of Texas Core Facility Award (RP170644). This study was supported by National Institutes of Health grants R01 GM083122 to D. Nicastro, R35 GM122574 to G.B. Witman, and R01 GM112050 to E.F. Smith, and by Cancer Prevention and Research Institute of Texas grant RR140082 to D. Nicastro. The authors declare no competing financial interests.
Publisher Copyright:
© 2019 Fu et al.
PY - 2019/12/2
Y1 - 2019/12/2
N2 - 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 Chlamydomonas 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.
AB - 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 Chlamydomonas 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.
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U2 - 10.1083/JCB.201906006
DO - 10.1083/JCB.201906006
M3 - Article
C2 - 31672705
AN - SCOPUS:85076062689
SN - 0021-9525
VL - 218
SP - 4236
EP - 4251
JO - Journal of Cell Biology
JF - Journal of Cell Biology
IS - 12
ER -