CMF70 is a subunit of the dynein regulatory complex.
Flagellar motility drives propulsion of several important pathogens and is essential for human development and physiology. Motility of the eukaryotic flagellum requires coordinate regulation of thousands of dynein motors arrayed along the axoneme, but the proteins underlying dynein regulation are largely unknown. The dynein regulatory complex, DRC, is recognized as a focal point of axonemal dynein regulation, but only a single DRC subunit, trypanin/PF2, is currently known. The component of motile flagella 70 protein, CMF70, is broadly and uniquely conserved among organisms with motile flagella, suggesting a role in axonemal motility. Here we demonstrate that CMF70 is part of the DRC from Trypanosoma brucei. CMF70 is located along the flagellum, co-sediments with trypanin in sucrose gradients and co-immunoprecipitates with trypanin. RNAi knockdown of CMF70 causes motility defects in a wild-type background and suppresses flagellar paralysis in cells with central pair defects, thus meeting the functional definition of a DRC subunit. Trypanin and CMF70 are mutually conserved in at least five of six extant eukaryotic clades, indicating that the DRC was probably present in the last common eukaryotic ancestor. We have identified only the second known subunit of this ubiquitous dynein regulatory system, highlighting the utility of combined genomic and functional analyses for identifying novel subunits of axonemal sub-complexes.
Kabututu
Zakayi P
ZP
Department of Microbiology, Immunology and Molecular Genetics, University of California, Los Angeles, CA 90095, USA.
Thayer
Michelle
M
Melehani
Jason H
JH
Hill
Kent L
KL
D000818:Animals; D056444:Axonemal Dyneins / Q000235:genetics / Q000378:metabolism; D005407:Flagella / Q000378:metabolism; D015151:Immunoblotting; D047468:Immunoprecipitation; D008854:Microscopy, Electron; D008856:Microscopy, Fluorescence; D015800:Protozoan Proteins / Q000235:genetics / Q000378:metabolism*; D034622:RNA Interference; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014346:Trypanosoma brucei brucei / Q000235:genetics / Q000378:metabolism*
D015800:Protozoan Proteins; C107172:T lymphocyte triggering factor, Trypanosoma brucei; D056444:Axonemal Dyneins