Calmodulin and PF6 are components of a complex that localizes to the C1 microtubule of the flagellar central apparatus.
Studies of flagellar motility in Chlamydomonas mutants lacking specific central apparatus components have supported the hypothesis that the inherent asymmetry of this structure provides important spatial cues for asymmetric regulation of dynein activity. These studies have also suggested that specific projections associated with the C1 and C2 central tubules make unique contributions to modulating motility; yet, we still do not know the identities of most polypeptides associated with the central tubules. To identify components of the C1a projection, we took an immunoprecipitation approach using antibodies generated against PF6. The pf6 mutant lacks the C1a projection and possesses flagella that only twitch; calcium-induced modulation of dynein activity on specific doublet microtubules is also defective in pf6 axonemes. Our antibodies specifically precipitated five polypeptides in addition to PF6. Using mass spectrometry, we determined the amino acid identities of these five polypeptides. Most notably, the PF6-containing complex includes calmodulin. Using antibodies generated against each precipitated polypeptide, we confirmed that these polypeptides comprise a single complex with PF6, and we identified specific binding partners for each member of the complex. The finding of a calmodulin-containing complex as an asymmetrically assembled component of the central apparatus implicates the central apparatus in calcium modulation of flagellar waveform.
Wargo
Matthew J
MJ
Dartmouth College, Department of Biological Sciences, 301 Gilman Hall, Hanover, NH 03755, USA.
Dymek
Erin E
EE
Smith
Elizabeth F
EF
D020418:Algal Proteins / Q000378:metabolism*; D000595:Amino Acid Sequence; D000818:Animals; D001692:Biological Transport; D002147:Calmodulin / Q000378:metabolism*; D002499:Centrifugation, Density Gradient; D016825:Chlamydomonas reinhardtii / Q000737:chemistry / Q000145:classification / Q000378:metabolism*; D005407:Flagella / Q000737:chemistry* / Q000502:physiology*; D013058:Mass Spectrometry; D008870:Microtubules / Q000378:metabolism*; D008969:Molecular Sequence Data; D010455:Peptides / Q000032:analysis; D011485:Protein Binding; D011993:Recombinant Fusion Proteins
D020418:Algal Proteins; D002147:Calmodulin; D010455:Peptides; D011993:Recombinant Fusion Proteins