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          Institute: MPI für molekulare Zellbiologie und Genetik     Collection: Publikationen mpi-cbg 2003     Display Documents



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ID: 14487.0, MPI für molekulare Zellbiologie und Genetik / Publikationen mpi-cbg 2003
Implication of a novel multiprotein Dam1p complex in outer kinetochore function
Authors:Cheeseman, I. M.; Brew, C.; Wolyniak, M.; Desai, A.; Anderson, S.; Muster, N.; Yates, J. R.; Huffaker, T. C.; Drubin, D. G.; Barnes, G.
Language:English
Date of Publication (YYYY-MM-DD):2001-12-24
Title of Journal:Journal of Cell Biology
Journal Abbrev.:J. Cell Biol.
Volume:155
Issue / Number:7
Start Page:1137
End Page:1145
Review Status:Peer-review
Audience:Not Specified
Abstract / Description:Dam1p, Duo1p, and Dad1p can associate with each other physically and are required for both spindle integrity and kinetochore function in budding yeast. Here, we present our purification from yeast extracts of an similar to 245 kD complex containing Dam1p, Duo1p, and Dad1p and Spc19p, Spc34p, and the previously uncharacterized proteins Dad2p and Ask1p. This Dam1p complex appears to be regulated through the phosphorylation of multiple subunits with at least one phosphorylation event changing during the cell cycle. We also find that purified Dam1p complex binds directly to microtubules in vitro with an affinity of similar to0.5 muM. To demonstrate that subunits of the Dam1p complex are functionally important for mitosis in vivo, we localized Spc19-green fluorescent protein (GFP), Spc34-GFP, Dad2-GFP, and Ask1-GFP to the mitotic spindle and to kinetochores and generated temperature-sensitive mutants of DAD2 and ASK1. These and other analyses implicate the four newly identified subunits and the Dam1p complex as a whole in outer kinetochore function where they are well positioned to facilitate the association of chromosomes with spindle microtubules.
Free Keywords:microtubule; spindle; mitosis; kinetochore; Saccharomyces cerevisiae
Comment of the Author/Creator:Date: 2001, DEC 24
External Publication Status:published
Document Type:Article
Communicated by:N. N.
Affiliations:MPI für molekulare Zellbiologie und Genetik
External Affiliations:Univ Calif Berkeley, Dept Cell & Mol Biol, 401 Barker Hall; 3202, Berkeley, CA 94720 USA; Univ Calif Berkeley, Dept Cell & Mol Biol, Berkeley, CA 94720 USA; Cornell Univ, Dept Mol Biol & Genet, Ithaca, NY 14853 USA; Scripps Res Ins, Dept Cell Biol, La Jolla, CA 92037 USA; Max Planck Inst Cell Biol & Genet, D-01307 Dresden, Germany
Identifiers:ISI:000173023500011 [ID No:1]
ISSN:0021-9525 [ID No:2]
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