Effects of HSP70 chaperones Ssa1 and Ssa2 on Ste5 scaffold and the mating mitogen-activated protein kinase (MAPK) pathway in Saccharomyces cerevisiae.
Effects of HSP70 chaperones Ssa1 and Ssa2 on Ste5 scaffold and the mating mitogen-activated protein kinase (MAPK) pathway in Saccharomyces cerevisiae.
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DOI：
10.1371/journal.pone.0289339
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发表时间：
2023
期刊：
影响因子：
3.7
通讯作者：
中科院分区：
文献类型：
--
作者：
Ste 5是所有真核生物中调节丝裂原活化蛋白激酶（MAPK）级联激活的支架蛋白的原型。Ste 5与许多蛋白质相关，包括Gβγ（Ste 4）、Ste 11 MAPKKK、Ste 7 MAPKK、Fus 3和Kss 1 MAPKs、Bem 1、Cdc 24。在这里，我们表明，Ste 5还与热休克蛋白70分子伴侣（Hsp 70）Ssa 1和Ssa 1和它的直系同源物Ssa 2是一起重要的Ste 5功能和有效的交配反应。大多数纯化过表达的Ste 5与Ssa 1相关。Ssa 1和Ssa 2的缺失对Ste 5丰度、完整性和定位具有有害影响，特别是当Ste 5以天然水平表达时。Ssa 1和Ssa 2的状态影响Ste 5电泳迁移率和被认为是磷酸化、泛素化和聚集的高分子量物质以及低分子量片段的形成。具有更大形成点状病灶倾向的Ste 5 VWA结构域突变体在突变位点附近结合Ssa 1的预测倾向降低，并且当Ssa 1过表达时形成更多点状病灶，支持Ste 5和Ssa 1之间的动态蛋白质质量控制关系。Ssa 1和Ssa 2的缺失降低了Fus 3和Kss 1 MAPK和FUS 1基因表达的激活，并损害了交配shmoo形态发生。令人惊讶的是，ssa 1，ssa 2，ssa 3和ssa 4单，双和三重突变体仍然可以交配，这表明存在补偿机制的折叠。进一步的分析表明，Ssa 1是交配和侵入性生长途径的主要Hsp 70分子伴侣，并揭示了交配形态发生所需的几个Hsp 70-Hsp 90分子伴侣网络蛋白。
Ste5 is a prototype of scaffold proteins that regulate activation of mitogen-activated protein kinase (MAPK) cascades in all eukaryotes. Ste5 associates with many proteins including Gβγ (Ste4), Ste11 MAPKKK, Ste7 MAPKK, Fus3 and Kss1 MAPKs, Bem1, Cdc24. Here we show that Ste5 also associates with heat shock protein 70 chaperone (Hsp70) Ssa1 and that Ssa1 and its ortholog Ssa2 are together important for Ste5 function and efficient mating responses. The majority of purified overexpressed Ste5 associates with Ssa1. Loss of Ssa1 and Ssa2 has deleterious effects on Ste5 abundance, integrity, and localization particularly when Ste5 is expressed at native levels. The status of Ssa1 and Ssa2 influences Ste5 electrophoresis mobility and formation of high molecular weight species thought to be phosphorylated, ubiquitinylated and aggregated and lower molecular weight fragments. A Ste5 VWA domain mutant with greater propensity to form punctate foci has reduced predicted propensity to bind Ssa1 near the mutation sites and forms more punctate foci when Ssa1 Is overexpressed, supporting a dynamic protein quality control relationship between Ste5 and Ssa1. Loss of Ssa1 and Ssa2 reduces activation of Fus3 and Kss1 MAPKs and FUS1 gene expression and impairs mating shmoo morphogenesis. Surprisingly, ssa1, ssa2, ssa3 and ssa4 single, double and triple mutants can still mate, suggesting compensatory mechanisms exist for folding. Additional analysis suggests Ssa1 is the major Hsp70 chaperone for the mating and invasive growth pathways and reveals several Hsp70-Hsp90 chaperone-network proteins required for mating morphogenesis.
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DOI：
10.1016/j.bbrc.2016.05.046
发表时间：
2016-06-17
影响因子：
3.1
作者：
Lee do H;Sherman MY;Goldberg AL
通讯作者：
Goldberg AL
DOI：
10.1073/pnas.88.21.9392
发表时间：
1991-11-01
影响因子：
11.1
作者：
ELION, EA;BRILL, JA;FINK, GR
通讯作者：
FINK, GR
DOI：
10.1074/jbc.r116.714980
发表时间：
2016-04-08
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
Alvaro CG;Thorner J
通讯作者：
Thorner J
影响因子：
8.8
作者：
Christiano R;Nagaraj N;Fröhlich F;Walther TC
通讯作者：
Walther TC
影响因子：
7.2
作者：
Chen, Bryan;Retzlaff, Marco;Frydman, Judith
通讯作者：
Frydman, Judith