Department of Molecular, Cellular, and Developmental Biology, University of Colorado, Boulder, CO 80309, USA.
Science. 2011 Jun 24;332(6037):1557-61. doi: 10.1126/science.1205193.
Centrosomes organize the bipolar mitotic spindle, and centrosomal defects cause chromosome instability. Protein phosphorylation modulates centrosome function, and we provide a comprehensive map of phosphorylation on intact yeast centrosomes (18 proteins). Mass spectrometry was used to identify 297 phosphorylation sites on centrosomes from different cell cycle stages. We observed different modes of phosphoregulation via specific protein kinases, phosphorylation site clustering, and conserved phosphorylated residues. Mutating all eight cyclin-dependent kinase (Cdk)-directed sites within the core component, Spc42, resulted in lethality and reduced centrosomal assembly. Alternatively, mutation of one conserved Cdk site within γ-tubulin (Tub4-S360D) caused mitotic delay and aberrant anaphase spindle elongation. Our work establishes the extent and complexity of this prominent posttranslational modification in centrosome biology and provides specific examples of phosphorylation control in centrosome function.
中心体组织双极有丝分裂纺锤体，中心体缺陷导致染色体不稳定。蛋白质磷酸化调节中心体的功能，我们提供了一个完整的酵母中心体磷酸化图谱（18 种蛋白质）。质谱法用于鉴定来自不同细胞周期阶段的中心体上的 297 个磷酸化位点。我们观察到通过特定蛋白激酶、磷酸化位点聚类和保守磷酸化残基的不同磷酸化调节模式。在核心成分 Spc42 内突变八个细胞周期蛋白依赖性激酶 (Cdk) 指导的位点会导致致死和中心体组装减少。相反，突变 γ-微管蛋白 (Tub4-S360D) 内的一个保守 Cdk 位点会导致有丝分裂延迟和异常的后期纺锤体伸长。我们的工作确定了这种在中心体生物学中突出的翻译后修饰的程度和复杂性，并提供了中心体功能中磷酸化控制的具体示例。
