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Structural organization of the C1a-e-c supercomplex within the ciliary central apparatus.

Gang Fu | Lei Zhao | Erin Dymek | Yuqing Hou | Kangkang Song | Nhan Phan | Zhiguo Shang | Elizabeth F Smith | George B Witman | Daniela Nicastro
The Journal of cell biology | 2019

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.

Pubmed ID: 31672705

Research resources used in this publication

None found

Antibodies used in this publication

None found

Associated grants

  • Agency: NIGMS NIH HHS, United States
    Id: R01 GM083122
  • Agency: NIGMS NIH HHS, United States
    Id: R01 GM112050
  • Agency: NIGMS NIH HHS, United States
    Id: R35 GM122574

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AxioVision Imaging System (tool)

RRID:SCR_002677

Digital image processing system where microscope settings and processing steps may be adjusted in single user interface. Can acquire images from variety of cameras. Includes software package for capturing, archiving and preparing images for publication. Allows users to visualize and present images in several dimensions. Functionality of imaging toolbox expands constantly with wide range of different modules that are tailored to specific applications or microscope accessories. This resource is duplicated by SCR_018376

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IMOD (tool)

RRID:SCR_003297

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Phytozome (tool)

RRID:SCR_006507

A comparative platform for green plant genomics. Families of orthologous and paralogous genes that represent the modern descendents of ancestral gene sets are constructed at key phylogenetic nodes. These families allow easy access to clade specific orthology / paralogy relationships as well as clade specific genes and gene expansions. As of release v9.1, Phytozome provides access to forty-one sequenced and annotated green plant genomes which have been clustered into gene families at 20 evolutionarily significant nodes. Where possible, each gene has been annotated with PFAM, KOG, KEGG, and PANTHER assignments, and publicly available annotations from RefSeq, UniProt, TAIR, JGI are hyper-linked and searchable.

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RRID:SCR_013318

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RRID:SCR_014960

Central repository that receives, catalogs, preserves, and distributes wild type and mutant cultures of the green alga Chlamydomonas reinhardtii, as well as useful molecular reagents and kits for education and research.

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nanoprobes.com (tool)

RRID:SCR_014323

A nanoparticle research collaborative which engineers nanoparticle products in order to find cures for cancer and other diseases. The gold labels produced are uncharged molecules which are cross-linked to specific sites on biomolecules. Labels can be attached to any molecule with a reactive group (proteins, peptides, oligonucleotides, small molecules and lipids) for detection and localization. New probes can be engineered based on any fragment of a naturally occurring biomolecule, and the label can be positioned away from the binding site so it does not interfere with binding.

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