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. 1978 Mar 1;76(3):729–747. doi: 10.1083/jcb.76.3.729

Chlamydomonas flagellar mutants lacking radial spokes and central tubules. Structure, composition, and function of specific axonemal components

PMCID: PMC2110011  PMID: 632325

Abstract

The fine structure, protein composition, and roles in flagellar movement of specific axonemal components were studied in wild-type Chlamydomonas and paralyzed mutants pf-14, pf-15A, and pf-19. Electron microscope examination of the isolated axoneme of pf-14 showed that it lacks the radial spokes but is otherwise structurally normal. Comparison of isolated axonemes of wild type and pf-14 by sodium dodecyl sulfate-acrylamide gel electrophoresis indicated that the mutant is missing a protein of 118,000 mol wt; this protein is apparently a major component of the spokes. Pf-15A and pf-19 lack the central tubules and sheath; axonemes of these mutants are missing three high molecular weight proteins which are probably components of the central tubule-central sheath complex. Under conditions where wild-type axonemes reactivated, axonemes of the three mutants remained intact but did not form bends. However, mutant and wild-type axonemes underwent identical adenosine triphosphate-induced disintegration after treatment with trypsin; the dynein arms of the mutants are therefore capable of generating interdoublet shearing forces. These findings indicated that both the radial spokes and the central tubule-central sheath complex are essential for conversion of interdoublet sliding into axonemal bending. Moreover, because axonemes of pf-14 remained intact under reactivating conditions, the nexin links alone are sufficient to limit the amount of interdoublet sliding that occurs. The axial periodicities of the central sheath, dynein arms, radial spokes, and nexin links of Chlamydomonas were determined by electron microscopy using the lattice- spacing of crystalline catalase as an internal standard. Some new ultrastructural details of the components are described.

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Selected References

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  1. AFZELIUS B. A. The fine structure of the cilia from ctenophore swimming-plates. J Biophys Biochem Cytol. 1961 Feb;9:383–394. doi: 10.1083/jcb.9.2.383. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Allen C., Borisy G. G. Structural polarity and directional growth of microtubules of Chlamydomonas flagella. J Mol Biol. 1974 Dec 5;90(2):381–402. doi: 10.1016/0022-2836(74)90381-7. [DOI] [PubMed] [Google Scholar]
  3. Allen R. D. A reinvestigation of cross-sections of cilia. J Cell Biol. 1968 Jun;37(3):825–831. doi: 10.1083/jcb.37.3.825. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Chasey D. Observations on the central pair of microtubules from the cilia of Tetrahymena pyriformis. J Cell Sci. 1969 Sep;5(2):453–458. doi: 10.1242/jcs.5.2.453. [DOI] [PubMed] [Google Scholar]
  5. Chasey D. The three-dimensional arrangement of radial spokes in the flagella of Chlamydomonas reinhardii. Exp Cell Res. 1974 Mar 15;84(1):374–380. doi: 10.1016/0014-4827(74)90418-2. [DOI] [PubMed] [Google Scholar]
  6. EBERSOLD W. T., LEVINE R. P., LEVINE E. E., OLMSTED M. A. Linkage maps in Chlamydomonas reinhardi. Genetics. 1962 May;47:531–543. doi: 10.1093/genetics/47.5.531. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. GIBBONS I. R. The relationship between the fine structure and direction of beat in gill cilia of a lamellibranch mollusc. J Biophys Biochem Cytol. 1961 Oct;11:179–205. doi: 10.1083/jcb.11.1.179. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Gibbons B. H., Gibbons I. R. The effect of partial extraction of dynein arms on the movement of reactivated sea-urchin sperm. J Cell Sci. 1973 Sep;13(2):337–357. doi: 10.1242/jcs.13.2.337. [DOI] [PubMed] [Google Scholar]
  9. Hopkins J. M. Subsidiary components of the flagella of Chlamydomonas reinhardii. J Cell Sci. 1970 Nov;7(3):823–839. doi: 10.1242/jcs.7.3.823. [DOI] [PubMed] [Google Scholar]
  10. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  11. LUFT J. H. Improvements in epoxy resin embedding methods. J Biophys Biochem Cytol. 1961 Feb;9:409–414. doi: 10.1083/jcb.9.2.409. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  13. Luftig R. An accurate measurement of the catalase crystal period and its use as an internal marker for electron microscopy. J Ultrastruct Res. 1967 Sep;20(1):91–102. doi: 10.1016/s0022-5320(67)80038-8. [DOI] [PubMed] [Google Scholar]
  14. Olson G. E., Linck R. W. Observations of the structural components of flagellar axonemes and central pair microtubules from rat sperm. J Ultrastruct Res. 1977 Oct;61(1):21–43. doi: 10.1016/s0022-5320(77)90004-1. [DOI] [PubMed] [Google Scholar]
  15. Piperno G., Huang B., Luck D. J. Two-dimensional analysis of flagellar proteins from wild-type and paralyzed mutants of Chlamydomonas reinhardtii. Proc Natl Acad Sci U S A. 1977 Apr;74(4):1600–1604. doi: 10.1073/pnas.74.4.1600. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. REYNOLDS E. S. The use of lead citrate at high pH as an electron-opaque stain in electron microscopy. J Cell Biol. 1963 Apr;17:208–212. doi: 10.1083/jcb.17.1.208. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Ringo D. L. Flagellar motion and fine structure of the flagellar apparatus in Chlamydomonas. J Cell Biol. 1967 Jun;33(3):543–571. doi: 10.1083/jcb.33.3.543. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Satir P. Studies on cilia. 3. Further studies on the cilium tip and a "sliding filament" model of ciliary motility. J Cell Biol. 1968 Oct;39(1):77–94. doi: 10.1083/jcb.39.1.77. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Summers K. E., Gibbons I. R. Adenosine triphosphate-induced sliding of tubules in trypsin-treated flagella of sea-urchin sperm. Proc Natl Acad Sci U S A. 1971 Dec;68(12):3092–3096. doi: 10.1073/pnas.68.12.3092. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Summers K. E., Gibbons I. R. Effects of trypsin digestion on flagellar structures and their relationship to motility. J Cell Biol. 1973 Sep;58(3):618–629. doi: 10.1083/jcb.58.3.618. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Thompson G. A., Jr, Baugh L. C., Walker L. F. Nonlethal deciliation of Tetrahymena by a local anesthetic and its utility as a tool for studying cilia regeneration. J Cell Biol. 1974 Apr;61(1):253–257. doi: 10.1083/jcb.61.1.253. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Warner F. D. Ciliary inter-microtubule bridges. J Cell Sci. 1976 Jan;20(1):101–114. doi: 10.1242/jcs.20.1.101. [DOI] [PubMed] [Google Scholar]
  23. Warner F. D., Satir P. The structural basis of ciliary bend formation. Radial spoke positional changes accompanying microtubule sliding. J Cell Biol. 1974 Oct;63(1):35–63. doi: 10.1083/jcb.63.1.35. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Witman G. B. The site of in vivo assembly of flagellar microtubules. Ann N Y Acad Sci. 1975 Jun 30;253:178–191. doi: 10.1111/j.1749-6632.1975.tb19199.x. [DOI] [PubMed] [Google Scholar]
  25. Wrigley N. G. The lattice spacing of crystalline catalase as an internal standard of length in electron microscopy. J Ultrastruct Res. 1968 Sep;24(5):454–464. doi: 10.1016/s0022-5320(68)80048-6. [DOI] [PubMed] [Google Scholar]

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