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J Cell Biol. 1993 February 1; 120(3): 733–741.
PMCID: PMC2119553

ptx1, a nonphototactic mutant of Chlamydomonas, lacks control of flagellar dominance

Abstract

A new mutant strain of Chlamydomonas, ptx1, has been identified which is defective in phototaxis. This strain swims with a rate and straightness of path comparable with that of wild-type cells, and retains the photoshock response. Thus, the mutation does not cause any gross defects in swimming ability or photoreception, and appears to be specific for phototaxis. Calcium is required for phototaxis in wild- type cells, and causes a concentration-dependent shift in flagellar dominance in reactivated, demembranated cell models. ptx1-reactivated models are defective in this calcium-dependent shift in flagellar dominance. This indicates that the mutation affects one or more components of the calcium-dependent axonemal regulatory system, and that this system mediates phototaxis. The reduction or absence of two 75-kD axonemal proteins correlates with the nonphototactic phenotype. Axonemal fractionation studies, and analysis of axonemes from mutant strains with known structural defects, failed to reveal the structural localization of the 75-kD proteins within the axoneme. The proteins are not components of the outer dynein arms, two of the three types of inner dynein arms, the radial spokes, or the central pair complex. Because changes in flagellar motility ultimately require the regulation of dynein activity, cell models from mutant strains defective in specific dynein arms were reactivated at various calcium concentrations. Mutants lacking the outer arms, or the I1 or I2 inner dynein arms, retain the wild-type calcium-dependent shift in flagellar dominance. Therefore, none of these arms are the sole mediators of phototaxis.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.
  • Bessen M, Fay RB, Witman GB. Calcium control of waveform in isolated flagellar axonemes of Chlamydomonas. J Cell Biol. 1980 Aug;86(2):446–455. [PMC free article] [PubMed]
  • Foster KW, Smyth RD. Light Antennas in phototactic algae. Microbiol Rev. 1980 Dec;44(4):572–630. [PMC free article] [PubMed]
  • Foster KW, Saranak J, Patel N, Zarilli G, Okabe M, Kline T, Nakanishi K. A rhodopsin is the functional photoreceptor for phototaxis in the unicellular eukaryote Chlamydomonas. Nature. 1984 Oct 25;311(5988):756–759. [PubMed]
  • Garrels JI. Two dimensional gel electrophoresis and computer analysis of proteins synthesized by clonal cell lines. J Biol Chem. 1979 Aug 25;254(16):7961–7977. [PubMed]
  • Gitelman SE, Witman GB. Purification of calmodulin from Chlamydomonas: calmodulin occurs in cell bodies and flagella. J Cell Biol. 1980 Dec;87(3 Pt 1):764–770. [PMC free article] [PubMed]
  • Hasegawa E, Hayashi H, Asakura S, Kamiya R. Stimulation of in vitro motility of Chlamydomonas axonemes by inhibition of cAMP-dependent phosphorylation. Cell Motil Cytoskeleton. 1987;8(4):302–311. [PubMed]
  • Hegemann P, Gärtner W, Uhl R. All-trans retinal constitutes the functional chromophore in Chlamydomonas rhodopsin. Biophys J. 1991 Dec;60(6):1477–1489. [PubMed]
  • Holmes JA, Dutcher SK. Cellular asymmetry in Chlamydomonas reinhardtii. J Cell Sci. 1989 Oct;94(Pt 2):273–285. [PubMed]
  • Huang B, Ramanis Z, Dutcher SK, Luck DJ. Uniflagellar mutants of Chlamydomonas: evidence for the role of basal bodies in transmission of positional information. Cell. 1982 Jul;29(3):745–753. [PubMed]
  • Huang B, Ramanis Z, Luck DJ. Suppressor mutations in Chlamydomonas reveal a regulatory mechanism for Flagellar function. Cell. 1982 Jan;28(1):115–124. [PubMed]
  • Kamiya R, Hasegawa E. Intrinsic difference in beat frequency between the two flagella of Chlamydomonas reinhardtii. Exp Cell Res. 1987 Nov;173(1):299–304. [PubMed]
  • Kamiya R, Okamoto M. A mutant of Chlamydomonas reinhardtii that lacks the flagellar outer dynein arm but can swim. J Cell Sci. 1985 Mar;74:181–191. [PubMed]
  • Kamiya R, Witman GB. Submicromolar levels of calcium control the balance of beating between the two flagella in demembranated models of Chlamydomonas. J Cell Biol. 1984 Jan;98(1):97–107. [PMC free article] [PubMed]
  • Kamiya R, Kurimoto E, Muto E. Two types of Chlamydomonas flagellar mutants missing different components of inner-arm dynein. J Cell Biol. 1991 Feb;112(3):441–447. [PMC free article] [PubMed]
  • King SM, Otter T, Witman GB. Purification and characterization of Chlamydomonas flagellar dyneins. Methods Enzymol. 1986;134:291–306. [PubMed]
  • King SM, Wilkerson CG, Witman GB. The Mr 78,000 intermediate chain of Chlamydomonas outer arm dynein interacts with alpha-tubulin in situ. J Biol Chem. 1991 May 5;266(13):8401–8407. [PubMed]
  • Melkonian M, Robenek H. Eyespot membranes of Chlamydomonas reinhardii: a freeze-fracture study. J Ultrastruct Res. 1980 Jul;72(1):90–102. [PubMed]
  • Morel-Laurens N. Calcium control of phototactic orientation in Chlamydomonas reinhardtii: sign and strength of response. Photochem Photobiol. 1987 Jan;45(1):119–128. [PubMed]
  • Morrissey JH. Silver stain for proteins in polyacrylamide gels: a modified procedure with enhanced uniform sensitivity. Anal Biochem. 1981 Nov 1;117(2):307–310. [PubMed]
  • Nultsch W. The photocontrol of movement of Chlamydomonas. Symp Soc Exp Biol. 1983;36:521–539. [PubMed]
  • O'Farrell PH. High resolution two-dimensional electrophoresis of proteins. J Biol Chem. 1975 May 25;250(10):4007–4021. [PMC free article] [PubMed]
  • Pfister KK, Fay RB, Witman GB. Purification and polypeptide composition of dynein ATPases from Chlamydomonas flagella. Cell Motil. 1982;2(6):525–547. [PubMed]
  • Piperno G, Luck DJ. Axonemal adenosine triphosphatases from flagella of Chlamydomonas reinhardtii. Purification of two dyneins. J Biol Chem. 1979 Apr 25;254(8):3084–3090. [PubMed]
  • Piperno G, Huang B, Ramanis Z, Luck DJ. Radial spokes of Chlamydomonas flagella: polypeptide composition and phosphorylation of stalk components. J Cell Biol. 1981 Jan;88(1):73–79. [PMC free article] [PubMed]
  • Stavis RL, Hirschberg R. Phototaxis in Chlamydomonas reinhardtii. J Cell Biol. 1973 Nov;59(2 Pt 1):367–377. [PMC free article] [PubMed]
  • Takahashi T, Yoshihara K, Watanabe M, Kubota M, Johnson R, Derguini F, Nakanishi K. Photoisomerization of retinal at 13-ene is important for phototaxis of Chlamydomonas reinhardtii: simultaneous measurements of phototactic and photophobic responses. Biochem Biophys Res Commun. 1991 Aug 15;178(3):1273–1279. [PubMed]
  • Witman GB. Isolation of Chlamydomonas flagella and flagellar axonemes. Methods Enzymol. 1986;134:280–290. [PubMed]
  • Witman GB, Plummer J, Sander G. Chlamydomonas flagellar mutants lacking radial spokes and central tubules. Structure, composition, and function of specific axonemal components. J Cell Biol. 1978 Mar;76(3):729–747. [PMC free article] [PubMed]

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