Phosphoproteome analysis of the human mitotic spindle
Phosphoproteome analysis of the human mitotic spindle
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DOI：
10.1073/pnas.0507066103
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发表时间：
2006-04-04
影响因子：
11.1
通讯作者：
Körner, R
中科院分区：
文献类型：
--
作者：
Nousiainen, M;Silljé, HHW;Körner, R
在细胞分裂期间，有丝分裂纺锤体将姐妹染色单体分离成两个新生细胞，使得每个子细胞继承一套完整的染色体。纺锤体形成错误可导致染色体错误分离和胞质分裂缺陷，从而导致基因组不稳定。为了确保纺锤体的正确功能，必须在时间和空间上严格调节纺锤体相关蛋白的活性和定位。可逆磷酸化已被证明是有丝分裂纺锤体组织的关键调节机制之一。然而，相对较低数量的确定在体内的磷酸化位点的纺锤体组件，阻碍了调控纺锤体网络的功能分析。因此，一个更完整的清单纺锤体相关蛋白的磷酸化位点将构成一个重要的进步。在这里，我们描述了基于质谱的鉴定纯化的人有丝分裂纺锤体的体内磷酸化位点。总共鉴定了736个磷酸化位点，其中312个可归因于已知的纺锤体蛋白。其中包括磷酸化位点，这些位点以前被证明对纺锤体相关蛋白的调节很重要。重要的是，该数据集还包括已知纺锤体蛋白的279个新的磷酸化位点，用于未来的功能研究。因此，纺锤体磷酸化位点的这一库存应作出重要贡献，以更好地了解调节有丝分裂纺锤体的形成，功能和完整性的分子机制。
During cell division, the mitotic spindle segregates the sister chromatids into two nascent cells, such that each daughter cell inherits one complete set of chromosomes. Errors in spindle formation can result in both chromosome missegregation and cytokinesis defects and hence lead to genomic instability. To ensure the correct function of the spindle, the activity and localization of spindle associated proteins has to be tightly regulated in time and space. Reversible phosphorylation has been shown to be one of the key regulatory mechanisms for the organization of the mitotic spindle. The relatively low number of identified in vivo phosphorylation sites of spindle components, however, has hampered functional analysis of regulatory spindle networks. A more complete inventory of the phosphorylation sites of spindle-associated proteins would therefore constitute an important advance. Here, we describe the mass spectrometry-based identification of in vivo phosphorylation sites from purified human mitotic spindles. In total, 736 phosphorylation sites were identified, of which 312 could be attributed to known spindle proteins. Among these are phosphorylation sites that were previously shown to be important for the regulation of spindle-associated proteins. Importantly, this data set also comprises 279 novel phosphorylation sites of known spindle proteins for future functional studies. This inventory of spindle phosphorylation sites should thus make an important contribution to a better understanding of the molecular mechanisms that regulate the formation, function, and integrity of the mitotic spindle.