Relating protein interaction networks to physiology by systematic mutant analyses

通过系统突变分析将蛋白质相互作用网络与生理学联系起来

基本信息

批准号：
10224929
负责人：
DANIEL N BOLON
金额：
$ 35.18万
依托单位：
UNIV OF MASSACHUSETTS MED SCH WORCESTER
依托单位国家：
美国
项目类别：
财政年份：
2015
资助国家：
美国
起止时间：
2015-02-01 至 2024-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10224929
关键词：
ATP phosphohydrolase Antineoplastic Agents Biochemical Biological Biophysics Calcineurin Catalytic Domain Cells Client Complex Dependence Development Disease Dominant-Negative Mutation Drug Targeting Drug resistance Engineering Fungal Drug Resistance Genes Glucocorticoid Receptor Grant Growth Health Human Individual Libraries Link Malignant Neoplasms Maps Medical Molecular Chaperones Mutation Mycoses Outcome Phosphotransferases Physiology Point Mutation Positioning Attribute Process Proteins Publishing Reporter Resolution Role Route Sampling Signal Transduction Signaling Protein Site Sodium Chloride Structure System Technology Therapeutic Variant Work Yeasts base biological adaptation to stress calcineurin phosphatase combat fitness fungus human disease improved inhibitor/antagonist insight mutant mutation screening next generation sequencing novel strategies pressure promoter protein protein interaction side effect tool transcription factor

项目摘要

Hsp90 is an essential eukaryotic chaperone that helps to produce and maintain the active state of a select set of substrates or clients that are disproportionately linked to signaling processes including many that promote cancer and the emergence of drug resistance in fungi. For these reasons, inhibitor strategies to manipulate the Hsp90 chaperone system promise many potential therapeutic benefits. Inhibitors targeting the ATPase site of Hsp90 effectively limit the emergence of drug resistance in fungi and show promise as anti-cancer agents. However, ATPase inhibitors block all known functions of Hsp90, leading to undesirable side effects. In our previous work, we generated a comprehensive library of all possible mutations at each of the 709 positions in Hsp90 and quantified the effects of each variant on yeast growth rate as a readout of overall network function integrated over all client proteins. We will build on this work to investigate Hsp90-client interaction mechanism. We will determine how four specific clients are impacted by Hsp90 mutations, which will identify sites on Hsp90 that could be targeted to inhibit specific clients. We will perform deep mutational scanning on specific client proteins to identify the biophysical features that determine Hsp90 dependence. In addition, we have developed a novel approach to quantify the dominant effects of Hsp90 mutations, which we are using as a powerful new route to understand its mechanism of action. Our studies will provide important mechanistic insights into a protein-protein interaction network that is biologically and medically important.

HSP90是必不可少的真核伴侣，有助于产生和维持活性状态一组选择与信号过程链接不成比例的基板或客户端包括许多促进癌症和真菌耐药性的出现的许多。为此原因，操纵HSP90伴侣系统的抑制剂策略承诺许多潜力治疗益处。针对HSP90 ATPase位点的抑制剂有效地限制了真菌中耐药性的出现，并表现出作为抗癌药物的希望。但是，ATPase抑制剂阻止了HSP90的所有已知功能，导致不良侧效果。在我们以前的工作中，我们在 HSP90中的709个位置中的每个位置，并量化了每个变体对酵母生长的影响速率作为整体网络功能的读数，这些函数在所有客户端蛋白质上集成。我们将基础这项研究用于研究HSP90客户相互作用机制。我们将确定如何四个特定客户受HSP90突变的影响，该突变将确定HSP90上的网站可能是针对抑制特定客户的目标。我们将对特定客户进行深入的突变扫描蛋白质鉴定决定HSP90依赖性的生物物理特征。另外，我们已经开发了一种新型方法来量化HSP90突变的主要作用，我们正在用作了解其行动机制的强大新途径。我们的研究将向蛋白质 - 蛋白质相互作用网络提供重要的机械洞察力生物学和医学上重要。