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

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

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cell Cycle Proteins / genetics
  • Cell Cycle Proteins / metabolism*
  • Chromosomal Proteins, Non-Histone / genetics
  • Chromosomal Proteins, Non-Histone / metabolism
  • Microtubules / metabolism
  • Mitosis
  • Multiprotein Complexes / genetics
  • Multiprotein Complexes / metabolism*
  • Mutagenesis, Site-Directed
  • Schizosaccharomyces / cytology
  • Schizosaccharomyces / genetics*
  • Schizosaccharomyces / metabolism
  • Schizosaccharomyces pombe Proteins / genetics
  • Schizosaccharomyces pombe Proteins / metabolism*
  • Spindle Apparatus / metabolism*

Substances

  • Cell Cycle Proteins
  • Chromosomal Proteins, Non-Histone
  • Cnp1 protein, S pombe
  • Mis19 protein, S pombe
  • Mis6 protein, S pombe
  • Multiprotein Complexes
  • Schizosaccharomyces pombe Proteins

Grants and funding

This work was supported by Grants-in-Aid for Young Scientists (A) (Grant Number 21687015) and for Scientific Research (B) (25291041) (to MS) and Grant-in-Aid for Scientific Research (S) (21227007) (to MY) from JSPS (http://www.jsps.go.jp/english/). This work was also supported by the Naito Foundation (https://www.naito-f.or.jp/en/), the Senri Life Science Foundation (http://www.senri-life.or.jp/), the Sumitomo Foundation (http://www.sumitomo.or.jp/e/) and Kato Life Science Foundation (https://www.katokinen.or.jp/) (to MS), and by Waseda University Grant for Special Research Projects (2013A-911, 2013B-178 and 2013A-6313 to MS.; 2013A-912 and 2013B-181 to KA). KA was a research fellow of JSPS. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.