Mitotic Spindle Integrity and Kinetochore Function Linked by the Duo1p/Dam1p Complex

doi:10.1083/jcb.152.1.197

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Published online 8 January 2001. doi:10.1083/jcb.152.1.197

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© The Rockefeller University Press, 0021-9525/2001/1/197/ $5.00
The Journal of Cell Biology, Volume 152, Number 1, January 8, 2001 197-212

Original Article

Mitotic Spindle Integrity and Kinetochore Function Linked by the Duo1p/Dam1p Complex

Iain M. Cheeseman^a, Maria Enquist-Newman^a, Thomas Müller-Reichert^a, David G. Drubin^a, and Georjana Barnes^a
^a Department of Molecular and Cell Biology, University of California at Berkeley, Berkeley, California 94720

Correspondence to: Georjana Barnes, 401 Barker Hall #3202, University of California at Berkeley, Berkeley, CA 94720-3202. Tel:(510) 642-5962 Fax:(510) 643-0062 E-mail:gbarnes{at}socrates.berkeley.edu.

Duo1p and Dam1p were previously identified as spindle proteinsin the budding yeast, Saccharomyces cerevisiae. Here, analysesof a diverse collection of duo1 and dam1 alleles were used todevelop a deeper understanding of the functions and interactionsof Duo1p and Dam1p. Based on the similarity of mutant phenotypes,genetic interactions between duo1 and dam1 alleles, interdependentlocalization to the mitotic spindle, and Duo1p/Dam1p coimmunoprecipitationfrom yeast protein extracts, these analyses indicated that Duo1pand Dam1p perform a shared function in vivo as components ofa protein complex. Duo1p and Dam1p are not required to assemblebipolar spindles, but they are required to maintain metaphaseand anaphase spindle integrity. Immunofluorescence and electronmicroscopy of duo1 and dam1 mutant spindles revealed a diversevariety of spindle defects. Our results also indicate a second,previously unidentified, role for the Duo1p/Dam1p complex. duo1and dam1 mutants show high rates of chromosome missegregation,premature anaphase events while arrested in metaphase, and geneticinteractions with a subset of kinetochore components consistentwith a role in kinetochore function. In addition, Duo1p andDam1p localize to kinetochores in chromosome spreads, suggestingthat this complex may serve as a link between the kinetochoreand the mitotic spindle.

Key Words: microtubule, spindle, mitosis, kinetochore, Saccharomyces cerevisiae

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