Proteomic analysis of in vivo 14-3-3 interactions in the yeast Saccharomyces cerevisiae
Proteomic analysis of in vivo 14-3-3 interactions in the yeast Saccharomyces cerevisiae
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DOI：
10.1021/bi700501t
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发表时间：
2007-07-03
期刊：
影响因子：
2.9
通讯作者：
Ichimura, Tohru
中科院分区：
文献类型：
--
作者：
Kakiuchi, Kazue;Yamauchi, Yoshio;Ichimura, Tohru
酿酒酵母产生两种14-3-3蛋白Bmh1和Bmh2，其确切功能尚不清楚。在这里，我们使用多步免疫亲和纯化和质谱进行了全面的蛋白质组学分析，并鉴定出271种酵母蛋白以磷酸化依赖的方式特异性结合Bmh1和-2。鉴定的蛋白质具有多种生化功能和细胞作用，包括细胞信号传导、代谢和细胞周期调节。重要的是，有许多蛋白质亚群通过各种细胞信号通路参与酵母生理学的调节，包括应激诱导的转录、细胞分裂和细胞壁的几丁质合成。事实上，我们发现酵母中Bmh1和-2缺失的突变体在MAPK （Hog1和Mpk1）级联的信号依赖性响应中存在缺陷，并且在芽颈处表现出异常的几丁质积累。我们认为，Bmh1和-2是许多细胞信号传导模块和蛋白质磷酸化介导的途径的共同调节剂，并通过协调控制已识别的多重磷酸化蛋白组分来调节各种生物事件。
The yeast Saccharomyces cerevisiae produces two 14-3-3 proteins, Bmh1 and Bmh2, whose exact functions have remained unclear. Here, we performed a comprehensive proteomic analysis using multistep immunoaffinity purification and mass spectrometry and identified 271 yeast proteins that specifically bind to Bmh1 and -2 in a phosphorylation-dependent manner. The identified proteins have diverse biochemical functions and cellular roles, including cell signaling, metabolism, and cell cycle regulation. Importantly, there are a number of protein subsets that are involved in the regulation of yeast physiology through a variety of cell signaling pathways, including stress-induced transcription, cell division, and chitin synthesis at the cell wall. In fact, we found that a yeast mutant deficient in Bmh1 and -2 had defects in signal-dependent response of the MAPK (Hog1 and Mpk1) cascade and exhibited an abnormal accumulation of chitin at the bud neck. We propose that Bmh1 and -2 are common regulators of many cell signaling modules and pathways mediated by protein phosphorylation and regulate a variety of biological events by coordinately controlling the identified multiplex phosphoprotein components.