Wong Jonathan, Nakajima Yuko, Westermann Stefan, Shang Ching, Kang Jung-Seog, Goodner Crystal, Houshmand Pantea, Fields Stanley, Chan Clarence S M, Drubin David, Barnes Georjana, Hazbun Tony
Department of Molecular and Cell Biology, University of California, Berkeley, CA 94720, USA.
Mol Biol Cell. 2007 Oct;18(10):3800-9. doi: 10.1091/mbc.e07-06-0536. Epub 2007 Jul 18.
The mitotic spindle consists of a complex network of proteins that segregates chromosomes in eukaryotes. To strengthen our understanding of the molecular composition, organization, and regulation of the mitotic spindle, we performed a system-wide two-hybrid screen on 94 proteins implicated in spindle function in Saccharomyces cerevisiae. We report 604 predominantly novel interactions that were detected in multiple screens, involving 303 distinct prey proteins. We uncovered a pattern of extensive interactions between spindle proteins reflecting the intricate organization of the spindle. Furthermore, we observed novel connections between kinetochore complexes and chromatin-modifying proteins and used phosphorylation site mutants of NDC80/TID3 to gain insights into possible phospho-regulation mechanisms. We also present analyses of She1p, a novel spindle protein that interacts with the Dam1 kinetochore/spindle complex. The wealth of protein interactions presented here highlights the extent to which mitotic spindle protein functions and regulation are integrated with each other and with other cellular activities.
有丝分裂纺锤体由一个复杂的蛋白质网络组成，该网络在真核生物中分离染色体。为了加强我们对有丝分裂纺锤体的分子组成、组织和调控的理解，我们对酿酒酵母中94种与纺锤体功能相关的蛋白质进行了全系统的双杂交筛选。我们报告了在多个筛选中检测到的604种主要为新的相互作用，涉及303种不同的猎物蛋白。我们发现了纺锤体蛋白之间广泛的相互作用模式，反映了纺锤体的复杂组织。此外，我们观察到动粒复合体与染色质修饰蛋白之间的新联系，并使用NDC80/TID3的磷酸化位点突变体来深入了解可能的磷酸化调控机制。我们还展示了对She1p的分析，She1p是一种与Dam1动粒/纺锤体复合体相互作用的新型纺锤体蛋白。这里呈现的丰富蛋白质相互作用突出了有丝分裂纺锤体蛋白功能和调控相互整合以及与其他细胞活动整合的程度。
