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dc.contributor.authorEcheverri, Christophe de Jesus
dc.contributor.authorPaschal, Bryce Mark
dc.contributor.authorVaughan, Kevin T.
dc.contributor.authorVallee, Richard B.
dc.date2022-08-11T08:08:57.000
dc.date.accessioned2022-08-23T16:13:38Z
dc.date.available2022-08-23T16:13:38Z
dc.date.issued1996-02-01
dc.date.submitted2008-09-04
dc.identifier.citation<p>J Cell Biol. 1996 Feb;132(4):617-33.</p>
dc.identifier.issn0021-9525 (Print)
dc.identifier.doi10.1083/jcb.132.4.617
dc.identifier.pmid8647893
dc.identifier.urihttp://hdl.handle.net/20.500.14038/33672
dc.description.abstractDynactin 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.
dc.language.isoen_US
dc.relation<p><a href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&list_uids=8647893&dopt=Abstract">Link to article in PubMed</a></p>
dc.relation.urlhttp://jcb.rupress.org/content/132/4/617.long
dc.subjectAmino 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
dc.subjectLife Sciences
dc.subjectMedicine and Health Sciences
dc.titleMolecular characterization of the 50-kD subunit of dynactin reveals function for the complex in chromosome alignment and spindle organization during mitosis
dc.typeJournal Article
dc.source.journaltitleThe Journal of cell biology
dc.source.volume132
dc.source.issue4
dc.identifier.legacyfulltexthttps://escholarship.umassmed.edu/cgi/viewcontent.cgi?article=1336&amp;context=gsbs_sp&amp;unstamped=1
dc.identifier.legacycoverpagehttps://escholarship.umassmed.edu/gsbs_sp/337
dc.identifier.contextkey619049
refterms.dateFOA2022-08-23T16:13:39Z
html.description.abstract<p>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.</p>
dc.identifier.submissionpathgsbs_sp/337
dc.contributor.departmentDepartment of Cell Biology
dc.contributor.departmentGraduate School of Biomedical Sciences
dc.source.pages617-33


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