Radial spokes of Chlamydomonas flagella: polypeptide composition and phosphorylation of stalk components

doi:10.1083/jcb.88.1.73

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Radial spokes of Chlamydomonas flagella: polypeptide composition and phosphorylation of stalk components

G Piperno, B Huang, Z Ramanis and DJ Luck

Polypeptides from flagella or axonemes of Chlamydomonas reinhardtii were analyzed by labeling cellular proteins by prolonged growth on 35S- containing media and using one- and two-dimensional electrophoretic techniques which can resolve greater than 170 axonemal components. By this approach, a paralyzed mutant that lacks axonemal radial spokes, pf14, has been shown to lack 17 polypeptides in the molecular weight range of 20,000 to 124,000 and in the isoelectric point range of 4.8- 7.1. Five of those polypeptides are also missing in the mutant pf-1 which lacks only radial spokeheads. The identification of the 17 polypeptides missing in pf-14 as components of radial spoke structures and the localization of the polypeptides lacking in pf-1 within the spokehead, are supported by experiments of chemical dissection of wild- type axonemes. Extraction procedures that solubilize outer and inner dynein arms preserve the structure of the radial spokes along with the 17 polypeptides in question. Six radial spoke polypeptides are solubilized in conditions that cause disassembly of radial spokeheads from the stalks and those components include the five polypeptides missing in pf-1. No Ca++- or Mg++-activated ATPase activities were found to be associated with solubilized preparations of wild-type radial spokeheads. In vivo pulse 32P incorporation experiments provide evidence that greater than 80 axonemal components are labeled by 32P and that five of the radial spoke stalk polypeptides are modified to different extents.
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Gingras, D., White, D., Garin, J., Cosson, J., Huitorel, P., Zingg, H., Cibert, C., Gagnon, C. (1998). Molecular Cloning and Characterization of a Radial Spoke Head Protein of Sea Urchin Sperm Axonemes: Involvement of the Protein in the Regulation of Sperm Motility. Mol. Biol. Cell 9: 513-522 [Abstract] [Full Text]
King, S. J., Dutcher, S. K. (1997). Phosphoregulation of an Inner Dynein Arm Complex in Chlamydomonas reinhardtii Is Altered in Phototactic Mutant Strains. JCB 136: 177-191 [Abstract] [Full Text]
Gingras, D., White, D., Garin, J., Multigner, L., Job, D., Cosson, J., Huitorel, P., Zingg, H., Dumas, F., Gagnon, C. (1996). Purification, Cloning, and Sequence Analysis of a Mr = 30,000 Protein from Sea Urchin Axonemes That Is Important for Sperm Motility. RELATIONSHIP OF THE PROTEIN TO A DYNEIN LIGHT CHAIN. J. Biol. Chem. 271: 12807-12813 [Abstract] [Full Text]
Smith, E., Sale, W. (1992). Regulation of dynein-driven microtubule sliding by the radial spokes in flagella. Science 257: 1557-1559 [Abstract]
Silflow, C. D., Lefebvre, P. A., McKeithan, T. W., Schloss, J. A., Keller, L. R., Rosenbaum, J. L. (1982). Expression of Flagellar Protein Genes during Flagellar Regeneration in Chlamydomonas. Cold Spring Harb Symp Quant Biol 46: 157-169 [Abstract]