Hirai Hayato, Arai Kunio, Kariyazono Ryo, Yamamoto Masayuki, Sato Masamitsu
Laboratory of Cytoskeletal Logistics, Department of Life Science and Medical Bioscience, Graduate School of Advanced Science and Technology, Waseda University, TWIns, Tokyo, Japan.
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.
PLoS One. 2014 Nov 6;9(11):e111905. doi: 10.1371/journal.pone.0111905. eCollection 2014.
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.
微管在广泛的细胞现象中发挥多种作用,包括细胞极性建立和染色体分离。已经鉴定出许多微管调节因子,包括微管相关蛋白和激酶,对这些因子的了解有助于我们从分子层面理解微管对每个相关细胞过程的调节。然而,已知的调节因子不足以解释这些过程是如何相互联系的,这突出了识别其他调节因子的必要性。为了找到这种新机制和微管调节因子,我们进行了一项筛选,将遗传学和显微镜技术结合起来,用于筛选裂殖酵母中微管组织有缺陷的突变体。我们分离出了大约900个在微管组织或核膜方面有缺陷的突变体,这些突变体被分为12类。我们特别关注一个突变体kis1,它在有丝分裂早期表现出纺锤体缺陷。kis1突变体经常无法组装正常的双极纺锤体。负责的基因编码一种着丝粒蛋白Mis19(也称为Eic1),它定位于着丝粒和纺锤体极的界面。我们还发现,在kis1细胞中,着丝粒内部蛋白Mis6/CENP-I和Cnp1/CENP-A从着丝粒上脱离,并且着丝粒与微管的附着存在缺陷。另一个突变体mis6也表现出类似的纺锤体缺陷。我们得出结论,Kis1是着丝粒内部组织所必需的,通过它Kis1确保着丝粒与微管的附着以及纺锤体的完整性。因此,我们提出了着丝粒内部组织与纺锤体完整性之间意想不到的关系。