Quantitative analysis of the evolving genotype-to-phenotype map

项目摘要 癌基因导向的癌症治疗和抗菌治疗经常受到快速的药物治疗的阻碍。 获得耐药性，持久缓解的主要障碍。有希望的洞察力正在显现 从一个看似遥远的领域-蛋白质折叠。为了发挥功能，蛋白质必须采用复杂的，通常 亚稳态构象危险的是，许多疾病都是由一个单一的蛋白质折叠或错误折叠引起的。 蛋白我以前的研究主要集中在Hsp 90，一种折叠的分子伴侣， 对致癌转化和信号传导至关重要的亚稳态蛋白质。通过影响褶皱 和功能的一个精英队列的调节蛋白，这种伴侣有能力影响 新特征的进化我将在我剩余的指导期间实现以下目标 培训和最初的独立研究生涯：（I）确定Hsp 90如何转化遗传 变化量Hsp 90强烈影响Mec 1/ATR多态性的作用，Mec 1/ATR是一个核心参与者。 在癌症信号传导中，以牺牲其他基因为代价，对某些基因毒性应激做出反应。 在生物化学和功能上，我将研究这如何影响DNA损伤反应，细胞， 周期和生存能力。此外，还将研究Hsp 90如何影响转录因子的转录。 Pdr 8是一种转录因子，能够对许多药物产生耐药性。最后要 研究Hsp 90抑制如何改变顺式调节基因多态性的影响， NDI 1的元素，以产生对氧化应激的强大抵抗力。(II)探讨 Hsp 90相关表型的同化。研究最终的突破性药物 抗性我将在最初和同化后分离出致病变异（当抗性被同化时）。 Hsp 90-独立的）。高通量遗传技术和下一代测序将 对这种现象提供了一种机械的理解。(III)识别和表征 其他因素，使高度突变的细胞生存，增殖，并演变新的 性状癌细胞必须承受大量的突变负荷， 不稳定我将筛选出能挽救高度突变菌株生长但不能 影响未变异的父母(IV)鉴定和表征其他基于蛋白质的 促进适应新环境的机制。利用数字核糖体 分析，酵母和细菌遗传学，生物化学我将描述两个朊病毒，[PSI+] 和[GAR+]，并确定它们如何影响适应不断变化的环境。的结果 这些研究将提供详细的了解如何热休克蛋白90，和蛋白质稳态更多 一般来说，控制信号通路，使耐药性和这些机制如何 有助于突破阻力。他们也会暴露出一个所有人都共有的致命弱点 癌症-对能够维持大量突变负荷的因素上瘾。

PROJECT SUMMARY Oncogene-directed cancer treatments and antimicrobial therapies are often thwarted by rapid acquisition of drug resistance, a chief barrier to lasting remission. Promising insight is emerging from a seemingly distant field - protein folding. To function, proteins must adopt complex, often metastable conformations. Perilously, many diseases arise from folding or misfolding of a single protein. My previous studies have focused on Hsp90, a molecular chaperone that folds metastable proteins critical for oncogenic transformation and signaling. By influencing the fold and function of an elite cohort of regulatory proteins, this chaperone has the power to influence the evolution of new traits. I will address the following aims during my remaining mentored training and initial independent research career: (I) Determine how Hsp90 transforms genetic variation. Hsp90 strongly impacts the effects of polymorphisms in Mec1/ATR, a central player in cancer signaling, enabling responses to certain genotoxic stresses at the expense of others. Biochemically and functionally, I will examine how this affects the DNA damage response, cell cycle, and viability. Additionally, is will investigate how Hsp90 impacts the transcriptional network of Pdr8, a transcription factor that enables resistance to many drugs. Finally, I will examine how Hsp90 inhibition transforms the effects of polymorphisms in cis-regulatory elements of NDI1, to create strong resistance to oxidative stresses. (II) Investigate assimilation of Hsp90-contingent phenotypes. To investigate eventual breakthrough drug resistance I will isolate causative variation initially and after assimilation (when resistance is Hsp90-independent). High-throughput genetic techniques and next-generation sequencing will provide a mechanistic understanding of this phenomenon. (III) Identify and characterize additional factors that allow highly mutated cells to survive, proliferate, and evolve new traits. Cancer cells must sustain massive mutation loads and consequently toxic proteome destabilization. I will screen for proteins that rescue growth of highly mutated strains but do not affect unmutated parents. (IV) Identify and characterize additional protein-based mechanisms that facilitate adaptation to new environments. Using digital ribosome profiling, yeast and bacterial genetics, and biochemistry I will characterize two prions, [PSI+] and [GAR+], and determine how they affect adaptation to changing environments. The results of these studies will offer detailed insight into how Hsp90, and protein homeostasis more generally, controls signaling pathways that enable drug resistance and how these mechanisms contribute to breakthrough resistance. They will also expose an Achilles' heel common to all cancers - addiction to factors that enable maintenance of massive mutation loads.