Fa1p is a 171 kDa protein essential for axonemal microtubule severing in Chlamydomonas

R. J. Finst, P. J. Kim, E. R. Griffis, L. M. Quarmby

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    Abstract

    A key event in deflagellation or deciliation is the severing of the nine outer-doublet axonemal microtubules at a specific site in the flagellar transition zone. Previous genetic analysis revealed three genes that are essential for deflagellation in Chlamydomonas. We have now identified the first of these products, Fa1p, a protein required for Ca(2+)-dependent, axonemal microtubule severing. Genetic mapping and the availability of a tagged allele allowed us to physically map the gene to the centromere-proximal domain of the mating-type locus. We identified clones of Chlamydomonas genomic DNA that rescued the Ca(2+)-dependent axonemal microtubule severing defect of fa1 mutants. The FA1 cDNA, obtained by RT-PCR, encodes a novel protein of 171 kDa, which is predicted to contain an amino-terminal coiled-coil domain and three Ca(2+)/calmodulin binding domains. By western analysis and subcellular fractionation, the FA1 product is enriched in flagellar-basal body complexes. Based on these observations and previous studies, we hypothesize that a Ca(2+)-activated, Ca(2+)-binding protein binds Fa1p leading ultimately to the activation of axonemal microtubule severing.
    Original languageEnglish
    Pages (from-to)1963-71
    Number of pages9
    JournalJournal of Cell Science
    Volume113
    Issue number11
    Publication statusPublished - 2000

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    Chlamydomonas
    Microtubules
    Proteins
    Basal Bodies
    Centromere
    Essential Genes
    Calmodulin
    Complementary DNA
    Clone Cells
    Alleles
    Polymerase Chain Reaction
    DNA
    Genes

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    Finst, R. J., Kim, P. J., Griffis, E. R., & Quarmby, L. M. (2000). Fa1p is a 171 kDa protein essential for axonemal microtubule severing in Chlamydomonas. Journal of Cell Science, 113(11), 1963-71.
    Finst, R. J. ; Kim, P. J. ; Griffis, E. R. ; Quarmby, L. M. / Fa1p is a 171 kDa protein essential for axonemal microtubule severing in Chlamydomonas. In: Journal of Cell Science. 2000 ; Vol. 113, No. 11. pp. 1963-71.
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    abstract = "A key event in deflagellation or deciliation is the severing of the nine outer-doublet axonemal microtubules at a specific site in the flagellar transition zone. Previous genetic analysis revealed three genes that are essential for deflagellation in Chlamydomonas. We have now identified the first of these products, Fa1p, a protein required for Ca(2+)-dependent, axonemal microtubule severing. Genetic mapping and the availability of a tagged allele allowed us to physically map the gene to the centromere-proximal domain of the mating-type locus. We identified clones of Chlamydomonas genomic DNA that rescued the Ca(2+)-dependent axonemal microtubule severing defect of fa1 mutants. The FA1 cDNA, obtained by RT-PCR, encodes a novel protein of 171 kDa, which is predicted to contain an amino-terminal coiled-coil domain and three Ca(2+)/calmodulin binding domains. By western analysis and subcellular fractionation, the FA1 product is enriched in flagellar-basal body complexes. Based on these observations and previous studies, we hypothesize that a Ca(2+)-activated, Ca(2+)-binding protein binds Fa1p leading ultimately to the activation of axonemal microtubule severing.",
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    Finst, RJ, Kim, PJ, Griffis, ER & Quarmby, LM 2000, 'Fa1p is a 171 kDa protein essential for axonemal microtubule severing in Chlamydomonas', Journal of Cell Science, vol. 113, no. 11, pp. 1963-71.

    Fa1p is a 171 kDa protein essential for axonemal microtubule severing in Chlamydomonas. / Finst, R. J.; Kim, P. J.; Griffis, E. R.; Quarmby, L. M.

    In: Journal of Cell Science, Vol. 113, No. 11, 2000, p. 1963-71.

    Research output: Contribution to journalArticle

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    AU - Kim, P. J.

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    AB - A key event in deflagellation or deciliation is the severing of the nine outer-doublet axonemal microtubules at a specific site in the flagellar transition zone. Previous genetic analysis revealed three genes that are essential for deflagellation in Chlamydomonas. We have now identified the first of these products, Fa1p, a protein required for Ca(2+)-dependent, axonemal microtubule severing. Genetic mapping and the availability of a tagged allele allowed us to physically map the gene to the centromere-proximal domain of the mating-type locus. We identified clones of Chlamydomonas genomic DNA that rescued the Ca(2+)-dependent axonemal microtubule severing defect of fa1 mutants. The FA1 cDNA, obtained by RT-PCR, encodes a novel protein of 171 kDa, which is predicted to contain an amino-terminal coiled-coil domain and three Ca(2+)/calmodulin binding domains. By western analysis and subcellular fractionation, the FA1 product is enriched in flagellar-basal body complexes. Based on these observations and previous studies, we hypothesize that a Ca(2+)-activated, Ca(2+)-binding protein binds Fa1p leading ultimately to the activation of axonemal microtubule severing.

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