A Comprehensive Membrane Interactome Mapping of Sho1p Reveals Fps1p as a Novel Key Player in the Regulation of the HOG Pathway in S. cerevisiae.
A Comprehensive Membrane Interactome Mapping of Sho1p Reveals Fps1p as a Novel Key Player in the Regulation of the HOG Pathway in S. cerevisiae.
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Sho1p 的综合膜相互作用组图谱揭示 Fps1p 作为酿酒酵母 HOG 途径调节中的新关键角色。
DOI：
10.1016/j.jmb.2015.01.016
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发表时间：
2015
影响因子：
5.6
通讯作者：
中科院分区：
文献类型：
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作者：
Lam,MandyHiuYi;Snider,Jamie;Rehal,Monique;Wong,Victoria;Aboualizadeh,Farzaneh;Drecun,Luka;Wong,Olivia;Jubran,Bellal;Li,Meirui;Ali,Mehrab;Jessulat,Matthew;Deineko,Viktor;Miller,Rachel;Lee,Mideum;Park,Hay-Oak;Davidson,Alan;
Sho1p是一种完整的膜蛋白，在酿酒酵母的高渗透压甘油(HOG)丝裂原活化蛋白激酶途径中起着至关重要的作用。在高渗透胁迫条件下被激活，它与其他HOG途径蛋白相互作用，介导细胞信号事件，确保酵母细胞能够适应并保持存活。为了进一步了解Sho1p的功能是如何通过其蛋白质-蛋白质相互作用(PPI)调节的，我们通过使用膜酵母双杂交(MYTH)鉴定了49个独特的Sho1p PPI，这是一种特别适合于在天然膜环境中鉴定全长完整膜蛋白的PPI的方法。通过文献搜索或两个互补的PPI分析进行二次验证，确认了80%的这些相互作用，从而产生了高质量的Sho1p互动组。这组推定的PPI包括先前描述的两个相互作用子，以及以前尚未被确定为与Sho1p结合的相互作用子集。Sho1p的SH3结构域被发现对与许多这些相互作用元件结合是重要的。一个特别感兴趣的新的相互作用因子是甘油转运蛋白Fps1p，它被证明需要Sho1p的SH3结构域通过其N端可溶的调节域结合。此外，我们还发现Fps1p参与了Sho1p功能的正向调节，并在下游激酶Hog1p的磷酸化中发挥作用。这项研究是迄今为止对Sho1p进行的最大规模的膜相互作用组分析，通过增加我们对Sho1p调控的理解，补充了过去对HOG途径的研究。
Sho1p, an integral membrane protein, plays a vital role in the high-osmolarity glycerol (HOG) mitogen-activated protein kinase pathway in the yeast Saccharomyces cerevisiae. Activated under conditions of high osmotic stress, it interacts with other HOG pathway proteins to mediate cell signaling events, ensuring that yeast cells can adapt and remain viable. In an attempt to further understand how the function of Sho1p is regulated through its protein–protein interactions (PPIs), we identified 49 unique Sho1p PPIs through the use of membrane yeast two-hybrid (MYTH), an assay specifically suited to identify PPIs of full-length integral membrane proteins in their native membrane environment. Secondary validation by literature search, or two complementary PPI assays, confirmed 80% of these interactions, resulting in a high-quality Sho1p interactome. This set of putative PPIs included both previously characterized interactors, along with a large subset of interactors that have not been previously identified as binding to Sho1p. The SH3 domain of Sho1p was found to be important for binding to many of these interactors. One particular novel interactor of interest is the glycerol transporter Fps1p, which was shown to require the SH3 domain of Sho1p for binding via its N-terminal soluble regulatory domain. Furthermore, we found that Fps1p is involved in the positive regulation of Sho1p function and plays a role in the phosphorylation of the downstream kinase Hog1p. This study represents the largest membrane interactome analysis of Sho1p to date and complements past studies on the HOG pathway by increasing our understanding of Sho1p regulation.