Multiple phosphorylation of the Cdc48/p97 cofactor protein Shp1/p47 occurs upon cell stress in budding yeast.
Multiple phosphorylation of the Cdc48/p97 cofactor protein Shp1/p47 occurs upon cell stress in budding yeast.
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DOI：
10.26508/lsa.202201642
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发表时间：
2023-04
影响因子：
4.4
通讯作者：
中科院分区：
文献类型：
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这项研究表明，磷酸化的酵母p97/Cdc 48衔接蛋白Shp 1增强后，多重应力，一个过程中调节的Mpk 1激酶和PP 1磷酸酶。在酵母中，由Cdc 48亚基组成的同六聚体p97复合物是包括ER相关降解在内的蛋白质质量控制途径的重要组成部分。该复合物以ATP依赖性方式分离蛋白质复合物，需要确定底物特异性的辅因子蛋白的参与。不同Cdc 48辅因子的功能以及它们如何被调节仍然相对知之甚少。在这项研究中，我们评估了Cdc 48衔接蛋白的磷酸化，揭示了一个独特的和独特的磷酸化模式的Shp 1/p47改变响应TORC 1抑制。定点突变证实，这种模式对应于磷酸化的残基S108和S315的Shp 1，与双磷酸化的形式成为主要的TORC 1抑制，ER应激和氧化应激。最后，我们评估了负责Shp 1磷酸化的候选激酶和磷酸酶，并确定了两种调节剂。我们发现，缺乏激酶Mpk 1/Slt 2的细胞显示Shp 1磷酸化减少，而受损的PP 1磷酸酶催化亚基（Glc 7）活性导致Shp 1磷酸化增加。总体而言，这些发现确定了磷酸化调节Shp 1在多个网站的Mpk 1激酶和PP 1磷酸酶后，各种压力。
This study shows that phosphorylation of the yeast p97/Cdc48 adaptor protein Shp1 is enhanced upon multiple stresses, a process regulated by both Mpk1 kinase and PP1 phosphatase. The homohexameric p97 complex, composed of Cdc48 subunits in yeast, is a crucial component of protein quality control pathways including ER-associated degradation. The complex acts to segregate protein complexes in an ATP-dependent manner, requiring the engagement of cofactor proteins that determine substrate specificity. The function of different Cdc48 cofactors and how they are regulated remains relatively poorly understood. In this study, we assess the phosphorylation of Cdc48 adaptor proteins, revealing a unique and distinctive phosphorylation pattern of Shp1/p47 that changed in response to TORC1 inhibition. Site-directed mutagenesis confirmed that this pattern corresponded to phosphorylation at residues S108 and S315 of Shp1, with the double-phosphorylated form becoming predominant upon TORC1 inhibition, ER-stress, and oxidative stress. Finally, we assessed candidate kinases and phosphatases responsible for Shp1 phosphorylation and identified two regulators. We found that cells lacking the kinase Mpk1/Slt2 show reduced Shp1 phosphorylation, whereas impaired PP1 phosphatase catalytic subunit (Glc7) activity resulted in increased Shp1 phosphorylation. Overall, these findings identify a phosphoregulation of Shp1 at multiple sites by Mpk1 kinase and PP1 phosphatase upon various stresses.
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DOI：
10.3390/jof8040368
发表时间：
2022-04-04
期刊：
Journal of fungi (Basel, Switzerland)
影响因子：
--
作者：
通讯作者：
--
影响因子：
21.3
作者：
Krastev DB;Li S;Sun Y;Wicks AJ;Hoslett G;Weekes D;Badder LM;Knight EG;Marlow R;Pardo MC;Yu L;Talele TT;Bartek J;Choudhary JS;Pommier Y;Pettitt SJ;Tutt ANJ;Ramadan K;Lord CJ
通讯作者：
Lord CJ
影响因子：
--
作者：
Pultz, Dennis;Bennetzen, Martin V.;Faergeman, Nils J.
通讯作者：
Faergeman, Nils J.
影响因子：
7
作者：
Kinoshita, E;Kinoshita-Kikuta, E;Koike, T
通讯作者：
Koike, T
影响因子：
4.8
作者：
Alberts, Sven M.;Sonntag, Caroline;Wolf, Dieter H.
通讯作者：
Wolf, Dieter H.