Haarer Brian K, Helfant Astrid Hoes, Nelson Scott A, Cooper John A, Amberg David C
Department of Biochemistry and Molecular Biology, SUNY Upstate Medical University, 750 E. Adams St., Syracuse, NY 13210, USA.
Eukaryot Cell. 2007 May;6(5):797-807. doi: 10.1128/EC.00332-06. Epub 2007 Apr 6.
Faithful partitioning of genetic material during cell division requires accurate spatial and temporal positioning of nuclei within dividing cells. In Saccharomyces cerevisiae, nuclear positioning is regulated by an elegant interplay between components of the actin and microtubule cytoskeletons. Regulators of this process include Bud6p (also referred to as the actin-interacting protein Aip3p) and Kar9p, which function to promote contacts between cytoplasmic microtubule ends and actin-delimited cortical attachment points. Here, we present the previously undetected association of Bud6p with the cytoplasmic face of yeast spindle pole bodies, the functional equivalent of metazoan centrosomes. Cells lacking Bud6p show exaggerated movements of the nucleus between mother and daughter cells and display reduced amounts of time a given spindle pole body spends in close association with the neck region of budding cells. Furthermore, overexpression of BUD6 greatly enhances interactions between the spindle pole body and mother-bud neck in a spindle alignment-defective dynactin mutant. These results suggest that association of either spindle pole body with neck components, rather than simply entry of a spindle pole body into the daughter cell, provides a positive signal for the progression of mitosis. We propose that Bud6p, through its localization at both spindle pole bodies and at the mother-bud neck, supports this positive signal and provides a regulatory mechanism to prevent excessive oscillations of preanaphase nuclei, thus reducing the likelihood of mitotic delays and nuclear missegregation.
在细胞分裂过程中,遗传物质的忠实分配需要细胞核在分裂细胞内进行精确的空间和时间定位。在酿酒酵母中,细胞核定位由肌动蛋白和微管细胞骨架成分之间的精妙相互作用调控。这个过程的调节因子包括Bud6p(也称为肌动蛋白相互作用蛋白Aip3p)和Kar9p,它们的作用是促进细胞质微管末端与肌动蛋白界定的皮质附着点之间的接触。在这里,我们展示了Bud6p与酵母纺锤体极体(后生动物中心体的功能等效物)的细胞质面之间以前未被检测到的关联。缺乏Bud6p的细胞在母细胞和子细胞之间显示出细胞核过度移动,并且给定的纺锤体极体与出芽细胞颈部紧密关联的时间减少。此外,BUD6的过表达在纺锤体排列缺陷的动力蛋白突变体中极大地增强了纺锤体极体与母芽颈部之间的相互作用。这些结果表明,纺锤体极体与颈部成分的关联,而不是简单地进入子细胞,为有丝分裂的进行提供了一个正向信号。我们提出,Bud6p通过其在纺锤体极体和母芽颈部的定位,支持这个正向信号,并提供一种调节机制来防止前期细胞核的过度振荡,从而降低有丝分裂延迟和核错误分离的可能性。