Quantitative analysis of the evolving genotype-to-phenotype map

项目总结 癌基因导向的癌症治疗和抗菌治疗经常受阻于快速 获得抗药性，这是持久缓解的主要障碍。前景光明的洞察力正在浮现 从一个看似遥远的领域--蛋白质折叠。为了发挥作用，蛋白质必须经常采用复杂的 亚稳构象。危险的是，许多疾病都是由一个折叠或错误折叠的单个 蛋白。我之前的研究重点是Hsp90，一种折叠的分子伴侣 对致癌转化和信号转导至关重要的亚稳蛋白。通过影响褶皱 和一组精英调控蛋白的功能，这种伴侣蛋白有能力影响 新特征的进化。在我剩下的指导期间，我将实现以下目标 培训和最初的独立研究生涯：(I)确定Hsp90如何转化基因 变种。HSP90强烈影响Mec1/ATR基因的多态效应 在癌症信号中，能够以牺牲其他人为代价对某些基因毒性应激做出反应。 从生物化学和功能上，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，IS将调查Hsp90如何影响转录 Pdr8是一种转录因子，可以对许多药物产生抗药性。最后，我会 研究Hsp90抑制如何改变顺式调控中的多态效应 NDI1的成分，对氧化应激产生强大的抵抗能力。(二)调查 Hsp90伴随表型的同化。研究最终的突破性药物 抗性I将在最初和同化后分离引起变异(当抗性是 不依赖于HSP90)。高通量基因技术和下一代测序将 提供对这一现象的机械理解。(三)确定和描述 允许高度突变的细胞存活、增殖并进化为新的细胞的其他因素 特征。癌细胞必须承受巨大的突变负荷，从而产生有毒的蛋白质组 不稳定。我会筛选能挽救高度突变菌株生长的蛋白质，但不能 影响未突变的双亲。(4)确定和表征其他以蛋白质为基础的 促进适应新环境的机制。使用数字核糖体 简写，酵母和细菌遗传学，以及生物化学I将表征两个Pron，[PSI+] 和[GAR+]，并确定它们如何影响对不断变化的环境的适应。结果是 这些研究将提供对Hsp90和蛋白质动态平衡更多的详细了解 通常情况下，控制导致耐药性的信号通路以及这些机制是如何 有助于突破阻力。他们还将暴露所有人共同的致命弱点 癌症--对能够维持大量突变负荷的因素的上瘾。

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.