Eukaryotic Cell
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EC Accepts, published online ahead of print on 27 June 2008
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Eukaryotic Cell doi:10.1128/EC.00138-08
Copyright (c) 2008, American Society for Microbiology and/or the Listed Authors/Institutions. All Rights Reserved.

The Microtubule Motor Protein Kar3 is Required for Normal Mitotic Division and Morphogenesis in Candida albicans

Racquel Kim Sherwood and Richard J. Bennett*

Department of Molecular Microbiology and Immunology, Brown University, Providence, RI 02912; Graduate Program in Molecular Biology, Cellular Biology, and Biochemistry, Brown University, Providence, RI 02912

* To whom correspondence should be addressed. Email: Richard_Bennett{at}brown.edu.


   Abstract

The kinesin-related protein Kar3 is a minus end directed molecular motor that plays a multifunctional role in microtubule-directed nuclear movement. Previously, it was shown that Candida albicans Kar3p is critical for nuclear fusion during mating as kar3 mutants were defective in karyogamy. In this study, we confirm that Kar3p is required for nuclear congression in mating, but that neither Kar3p nor the dynein motor protein Dyn1p are required for nuclear migration in the mating projection prior to cell fusion. In addition, we show that C. albicans Kar3p plays an important role in the cell and colony morphology of mitotically dividing cells, as evidenced by diminished filamentation of kar3 cells on Spider medium and an increased tendency of mutant cells to form pseudohyphal cells in liquid culture. Loss of Kar3p also led to defects in nuclear division, causing cells to grow slowly and exhibit reduced viability compared to wildtype cells. Slow growth was due, at least in part, to delayed cell cycle progression, as cells lacking Kar3p accumulated in anaphase of the cell cycle. Consistent with a role in mitotic division, Kar3 protein was shown to localize to the spindle pole bodies. Finally, kar3 cells exhibited unstable or aberrant mitotic spindles, a finding that accounts for the delay in cell cycle progression and decreased viability of these cells. We suggest that the altered morphology of kar3 cells is a direct consequence of the delay in anaphase, and this leads to increased polarized growth and pseudohyphae formation.







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