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    The 78,000 M(r) intermediate chain of Chlamydomonas outer arm dynein isa WD-repeat protein required for arm assembly

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    Authors
    Wilkerson, Curtis G.
    King, Stephen M.
    Koutoulis, Anthony
    Pazour, Gregory J.
    Witman, George B.
    UMass Chan Affiliations
    Program in Molecular Medicine
    Department of Cell Biology
    Document Type
    Journal Article
    Publication Date
    1995-04-01
    Keywords
    Amino Acid Sequence
    Animals
    Base Sequence
    Blotting, Southern
    Chlamydomonas reinhardtii
    Consensus Sequence
    Dynein ATPase
    Flagella
    Macromolecular Substances
    Microtubules
    Molecular Sequence Data
    Molecular Weight
    Mutagenesis, Insertional
    Oligonucleotide Probes
    Protein Biosynthesis
    Recombinant Proteins
    *Repetitive Sequences, Nucleic Acid
    Sequence Homology, Amino Acid
    Transcription, Genetic
    Amino Acids, Peptides, and Proteins
    Cell Biology
    Genetic Phenomena
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    Link to Full Text
    http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2120364
    Abstract
    We have isolated and sequenced a full-length cDNA clone encoding the 78,000 Mr intermediate chain (IC78) of the Chlamydomonas outer arm dynein. This protein previously was shown to be located at the base of the solubilized dynein particle and to interact with alpha tubulin in situ, suggesting that it may be involved in binding the outer arm to the doublet microtubule. The sequence predicts a polypeptide of 683 amino acids having a mass of 76.5 kD. Sequence comparison indicates that IC78 is homologous to the 69,000 M(r) intermediate chain (IC69) of Chlamydomonas outer arm dynein and to the 74,000 M(r) intermediate chain (IC74) of cytoplasmic dynein. The similarity between the chains is greatest in their COOH-terminal halves; the NH(2)-terminal halves are highly divergent. The COOH-terminal half of IC78 contains six short imperfect repeats, termed WD repeats, that are thought to be involved in protein-protein interactions. Although not previously reported, these repeated elements also are present in IC69 and IC74. Using the IC78 cDNA as a probe, we screened a group of slow-swimming insertional mutants and identified one which has a large insertion in the IC78 gene and seven in which the IC78 gene is completely deleted. Electron microscopy of three of these IC78 mutants revealed that each is missing the outer arm, indicating that IC78 is essential for arm assembly or attachment to the outer doublet. Restriction fragment length polymorphism mapping places the IC78 gene on the left arm of chromosome XII/XIII, at or near the mutation oda9, which also causes loss of the outer arm. Mutants with defects in the IC78 gene do not complement the oda9 mutation in stable diploids, strongly suggesting that ODA9 is the structural gene for IC78.
    Source

    J Cell Biol. 1995 Apr;129(1):169-78.

    DOI
    10.1083/jcb.129.1.169
    Permanent Link to this Item
    http://hdl.handle.net/20.500.14038/26418
    PubMed ID
    7698982
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    Link to Article in PubMed

    ae974a485f413a2113503eed53cd6c53
    10.1083/jcb.129.1.169
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