Analyses of functional domains within the PF6 protein of the central apparatus reveal a role for PF6 sub-complex members in regulating flagellar beat frequency

Numerous studies have indicated that each of the seven projections associated with the central pair of microtubules plays a distinct role in regulating eukaryotic ciliary/flagellar motility. Mutants which lack specific projections have distinct motility phenotypes. For example, Chlamydomonas pf6 mutants lack the C1a projection and have twitchy, non-beating flagella. The C1a projection is a complex of proteins including PF6, C1a-86, C1a-34, C1a-32, C1a-18, and calmodulin. To define functional domains within PF6 and to potentially assign functions to specific C1a components, we generated deletion constructs of the PF6 gene and tested for their ability to assemble and rescue motility upon transformation of mutant pf6 cells. Our results demonstrate that domains near the carboxyl-terminus of PF6 are essential for motility and/or assembly of the projection. The amino terminal half of PF6 is not required for C1a assembly; however, this region is important for stability of the C1a-34, C1a-32, and C1a-18 sub-complex and wild-type beat frequency. Analysis of double mutants lacking the amino terminus of PF6 and outer dynein arms reveal that C1a may play a role in modulating both inner and outer dynein arm activity. (C) 2012 Wiley Periodicals, Inc