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    Molecular characterization of the 50-kD subunit of dynactin reveals function for the complex in chromosome alignment and spindle organization during mitosis

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    Authors
    Echeverri, Christophe de Jesus
    Paschal, Bryce Mark
    Vaughan, Kevin T.
    Vallee, Richard B.
    UMass Chan Affiliations
    Department of Cell Biology
    Graduate School of Biomedical Sciences
    Document Type
    Journal Article
    Publication Date
    1996-02-01
    Keywords
    Amino Acid Sequence; Animals; Base Sequence; Cattle; Cell Line; *Chromosomes; Cloning, Molecular; DNA, Complementary; Dynein ATPase; Humans; Male; Microtubule Proteins; *Microtubule-Associated Proteins; *Mitosis; Molecular Sequence Data; Rats; Rats, Sprague-Dawley; Sequence Analysis
    Life Sciences
    Medicine and Health Sciences
    
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    Link to Full Text
    http://jcb.rupress.org/content/132/4/617.long
    Abstract
    Dynactin is a multi-subunit complex which has been implicated in cytoplasmic dynein function, though its mechanism of action is unknown. In this study, we have characterized the 50-kD subunit of dynactin, and analyzed the effects of its overexpression on mitosis in living cells. Rat and human cDNA clones revealed p50 to be novel and highly conserved, containing three predicted coiled-coil domains. Immunofluorescence staining of dynactin and cytoplasmic dynein components in cultured vertebrate cells showed that both complexes are recruited to kinetochores during prometaphase, and concentrate near spindle poles thereafter. Overexpression of p50 in COS-7 cells disrupted mitosis, causing cells to accumulate in a prometaphase-like state. Chromosomes were condensed but unaligned, and spindles, while still bipolar, were dramatically distorted. Sedimentation analysis revealed the dynactin complex to be dissociated in the transfected cultures. Furthermore, both dynactin and cytoplasmic dynein staining at prometaphase kinetochores was markedly diminished in cells expressing high levels of p50. These findings represent clear evidence for dynactin and cytoplasmic dynein codistribution within cells, and for the presence of dynactin at kinetochores. The data also provide direct in vivo evidence for a role for vertebrate dynactin in modulating cytoplasmic dynein binding to an organelle, and implicate both dynactin and dynein in chromosome alignment and spindle organization.
    Source

    J Cell Biol. 1996 Feb;132(4):617-33.

    DOI
    10.1083/jcb.132.4.617
    Permanent Link to this Item
    http://hdl.handle.net/20.500.14038/33672
    PubMed ID
    8647893
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    Link to article in PubMed

    ae974a485f413a2113503eed53cd6c53
    10.1083/jcb.132.4.617
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