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PLoS One. 2014 Nov 6;9(11):e111905. doi: 10.1371/journal.pone.0111905. eCollection 2014.

The kinetochore protein Kis1/Eic1/Mis19 ensures the integrity of mitotic spindles through maintenance of kinetochore factors Mis6/CENP-I and CENP-A.

Author information

1
Laboratory of Cytoskeletal Logistics, Department of Life Science and Medical Bioscience, Graduate School of Advanced Science and Technology, Waseda University, TWIns, Tokyo, Japan.
2
Laboratory of Cytoskeletal Logistics, Department of Life Science and Medical Bioscience, Graduate School of Advanced Science and Technology, Waseda University, TWIns, Tokyo, Japan; Department of Biophysics and Biochemistry, Graduate School of Science, University of Tokyo, Tokyo, Japan.
3
Department of Biophysics and Biochemistry, Graduate School of Science, University of Tokyo, Tokyo, Japan.
4
Department of Biophysics and Biochemistry, Graduate School of Science, University of Tokyo, Tokyo, Japan; Laboratory of Cell Responses, National Institute for Basic Biology, Aichi, Japan.
5
Laboratory of Cytoskeletal Logistics, Department of Life Science and Medical Bioscience, Graduate School of Advanced Science and Technology, Waseda University, TWIns, Tokyo, Japan; Department of Biophysics and Biochemistry, Graduate School of Science, University of Tokyo, Tokyo, Japan; PRESTO, Japan Science and Technology Agency, Saitama, Japan.

Abstract

Microtubules play multiple roles in a wide range of cellular phenomena, including cell polarity establishment and chromosome segregation. A number of microtubule regulators have been identified, including microtubule-associated proteins and kinases, and knowledge of these factors has contributed to our molecular understanding of microtubule regulation of each relevant cellular process. The known regulators, however, are insufficient to explain how those processes are linked to one another, underscoring the need to identify additional regulators. To find such novel mechanisms and microtubule regulators, we performed a screen that combined genetics and microscopy for fission yeast mutants defective in microtubule organization. We isolated approximately 900 mutants showing defects in either microtubule organization or the nuclear envelope, and these mutants were classified into 12 categories. We particularly focused on one mutant, kis1, which displayed spindle defects in early mitosis. The kis1 mutant frequently failed to assemble a normal bipolar spindle. The responsible gene encoded a kinetochore protein, Mis19 (also known as Eic1), which localized to the interface of kinetochores and spindle poles. We also found that the inner kinetochore proteins Mis6/CENP-I and Cnp1/CENP-A were delocalized from kinetochores in the kis1 cells and that kinetochore-microtubule attachment was defective. Another mutant, mis6, also displayed similar spindle defects. We conclude that Kis1 is required for inner kinetochore organization, through which Kis1 ensures kinetochore-microtubule attachment and spindle integrity. Thus, we propose an unexpected relationship between inner kinetochore organization and spindle integrity.

PMID:
25375240
PMCID:
PMC4222959
DOI:
10.1371/journal.pone.0111905
[Indexed for MEDLINE]
Free PMC Article

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