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Vol. 12, Issue 3, 739-751, March 2001

The Chlamydomonas PF6 Locus Encodes a Large Alanine/Proline-Rich Polypeptide That Is Required for Assembly of a Central Pair Projection and Regulates Flagellar Motility

Gerald Rupp,*dagger Eileen O'Toole,Dagger and Mary E. Porter*§

 *Department of Genetics, Cell Biology, and Development, University of Minnesota Medical School, Minneapolis, Minnesota 55455; and  Dagger Department of Molecular, Cellular, and Developmental Biology, University of Colorado at Boulder, Boulder, Colorado 80309

Efficient motility of the eukaryotic flagellum requires precise temporal and spatial control of its constituent dynein motors. The central pair and its associated structures have been implicated as important members of a signal transduction cascade that ultimately regulates dynein arm activity. To identify central pair components involved in this process, we characterized a Chlamydomonas motility mutant (pf6-2) obtained by insertional mutagenesis. pf6-2 flagella twitch ineffectively and lack the 1a projection on the C1 microtubule of the central pair. Transformation with constructs containing a full-length, wild-type copy of the PF6 gene rescues the functional, structural, and biochemical defects associated with the pf6 mutation. Sequence analysis indicates that the PF6 gene encodes a large polypeptide that contains numerous alanine-rich, proline-rich, and basic domains and has limited homology to an expressed sequence tag derived from a human testis cDNA library. Biochemical analysis of an epitope-tagged PF6 construct demonstrates that the PF6 polypeptide is an axonemal component that cosediments at 12.6S with several other polypeptides. The PF6 protein appears to be an essential component required for assembly of some of these polypeptides into the C1-1a projection.

dagger Present address: Department of Anatomy, Southern Illinois University School of Medicine, Carbondale, IL 62901.

§ Corresponding author. E-mail address: mary-p{at}biosci.cbs.umn.edu.

Molecular Biology of the Cell
Vol. 12, 739-751, March 2001
Copyright © 2001 by The American Society for Cell Biology


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