Quantitative proteomic analysis of yeast DNA replication proteins
Quantitative proteomic analysis of yeast DNA replication proteins
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DOI：
10.1016/j.ymeth.2012.03.012
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发表时间：
2012-06-01
期刊：
影响因子：
4.8
通讯作者：
Donaldson, Anne D.
中科院分区：
文献类型：
--
作者：
Kubota, Takashi;Stead, David A.;Donaldson, Anne D.
染色质是动态调节的，对其组成的蛋白质组学分析可以提供有关染色质功能成分的重要信息。例如，许多DNA复制蛋白在细胞周期的特定时间与染色质结合。蛋白质组学研究也可以用来表征染色质组成在DNA损伤等扰动下的变化，而有用的信息是通过测试染色体稳定途径中突变对染色质组成的影响来获得的。我们已经成功地利用细胞培养中氨基酸稳定同位素标记(SILAC)的方法对酵母染色质的正常和病理变化进行了蛋白质组学定量分析。在这里，我们描述了这种蛋白质组学方法来分析酿酒酵母染色质的变化，并通过分析细胞从G1期(由阿尔法因子诱导)到S期(在DNA复制抑制剂羟基脲的存在下)染色质组成的变化来说明这一过程。(C)2012 Elsevier Inc.保留所有权利。
Chromatin is dynamically regulated, and proteomic analysis of its composition can provide important information about chromatin functional components. Many DNA replication proteins for example bind chromatin at specific times during the cell cycle. Proteomic investigation can also be used to characterize changes in chromatin composition in response to perturbations such as DNA damage, while useful information is obtained by testing the effects on chromatin composition of mutations in chromosome stability pathways. We have successfully used the method of stable isotope labeling by amino acids in cell culture (SILAC) for quantitative proteomic analysis of normal and pathological changes to yeast chromatin. Here we describe this proteomic method for analyzing changes to Saccharomyces cerevisiae chromatin, illustrating the procedure with an analysis of the changes that occur in chromatin composition as cells progress from a G1 phase block (induced by alpha factor) into S phase (in the presence of DNA replication inhibitor hydroxyurea). (c) 2012 Elsevier Inc. All rights reserved.