Systematic Mutant Analyses Elucidate General and Client-Specific Aspects of Hsp90 Function.
Systematic Mutant Analyses Elucidate General and Client-Specific Aspects of Hsp90 Function.
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DOI：
10.1016/j.celrep.2016.03.046
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发表时间：
2016-04-19
期刊：
影响因子：
8.8
通讯作者：
Bolon DNA
中科院分区：
文献类型：
--
作者：
Mishra P;Flynn JM;Starr TN;Bolon DNA
为了探测真核生物中许多信号蛋白成熟所必需的HSP90伴侣的机制，我们分析了ATPase结构域中所有单个氨基酸变化对酵母生长速率的影响。对突变的位置的敏感性受到与ATP磷酸盐的接近性的强烈影响，表明ATPase驱动的构象变化对HSP90施加了严格的物理约束。为了调查这些约束对于不同客户的变化，我们对跨越观察到的适应性效应的全范围的HSP90突变体进行了生化分析。我们观察到九个HSP90突变对V-SRC和糖皮质激素受体（GR）激活的明显影响，表明可以为这些客户使用不同的伴侣机制。这些结果为理解HSP90机制提供了详细的指南，并突出了针对客户子集的HSP90抑制剂的潜力。
To probe the mechanism of the Hsp90 chaperone that is required for the maturation of many signaling proteins in eukaryotes, we analyzed the effects of all individual amino acid changes in the ATPase domain on yeast growth rate. The sensitivity of a position to mutation was strongly influenced by proximity to the phosphates of ATP, indicating that ATPase driven conformational changes impose stringent physical constraints on Hsp90. To investigate how these constraints may vary for different clients, we performed biochemical analyses on a panel of Hsp90 mutants spanning the full range of observed fitness effects. We observed distinct effects of nine Hsp90 mutations on activation of v-src and glucocorticoid receptor (GR), indicating that different chaperone mechanisms can be utilized for these clients. These results provide a detailed guide for understanding Hsp90 mechanism and highlight the potential for inhibitors of Hsp90 that target a subset of clients.