Quantitative analysis of the evolving genotype-to-phenotype map

不断演变的基因型到表型图谱的定量分析

• 批准号: 8600700
• 负责人: Daniel Jarosz
• 金额: $ 24.27万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-07-01 至 2015-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8600700
• 关键词: Address; Adopted; Affect; Assimilations; Bacteria; Biochemistry; Carbon; Cell Cycle; Cells; Chemicals; Clinical; Collaborations; Communication; Complex; DNA Damage; DNA damage checkpoint; Dependence; Disease; Disease remission; Distant; Drug resistance; Enabling Factors; Environment; Evolution; Fungal Drug Resistance; Genes; Genetic; Genetic Polymorphism; Genetic Techniques; Genetic Variation; Genotoxic Stress; Genotype; Growth; Homeostasis; Human Pathology; Infection; Institutes; Maintenance; Malignant Neoplasms; Maps; Mentors; Messenger RNA; Molecular; Molecular Chaperones; Molecular Conformation; Multi-Drug Resistance; Mutate; Mutation; NADH dehydrogenase (ubiquinone); Nerve Degeneration; Oncogenes; Oncogenic; Oxidative Stress; Parents; Pharmaceutical Preparations; Phenotype; Phosphotransferases; Prions; Process; Proliferating; Protein Conformation; Protein Structure Initiative; Proteins; Proteome; Quantitative Genetics; Regulatory Element; Research; Resistance; Ribosomes; Saccharomyces cerevisiae; Sentinel; Shapes; Signal Pathway; Signal Transduction; Source; Stress; System; Techniques; Therapeutic Intervention; Toxic effect; Training; Variant; Yeasts; antimicrobial; bacterial genetics; base; cancer addiction; cancer cell; cancer therapy; career; cohort; digital; environmental change; genetic regulatory protein; genome wide association study; insight; next generation sequencing; novel; protein folding; protein function; reconstruction; resistance mechanism; response; trait; transcription factor; yeast genetics

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.

项目摘要 致癌基因指导的癌症治疗和抗菌疗法通常会因快速而挫败 获得耐药性，这是持久缓解的主要障碍。有希望的见识正在出现 从看似遥远的场 - 蛋白质折叠。为了起作用，蛋白质必须采用复杂的，通常 亚稳态构象。危险的是，许多疾病是由于单一的折叠或错误折叠而引起的 蛋白质。我以前的研究集中于HSP90，这是一种折叠的分子伴侣 亚稳态蛋白对于致癌转化和信号传导至关重要。通过影响折叠 和精英群体的调节蛋白的功能，该伴侣有能力影响 新特征的演变。在剩下的指导期间，我将解决以下目标 培训和初始独立研究职业：（i）确定HSP90如何改变遗传 变化。 HSP90强烈影响多态性在MEC1/ATR中的影响 在癌症信号传导中，以牺牲他人为代价的某些遗传毒性应激的反应。 在生化和功能上，我将检查这如何影响DNA损伤响应，细胞 周期和生存能力。此外，将研究HSP90如何影响转录 PDR8网络，这是一种转录因子，可抗许多药物。最后，我会的 检查HSP90抑制如何改变多态性在顺式调节中的影响 NDI1的元素，以对氧化应激产生强烈的抗性。 （ii）调查 HSP90换型表型的同化。调查最终突破性药物 抗性i最初和同化后会分离病因变异（当电阻为 HSP90独立于）。高通量遗传技术和下一代测序将 提供对这一现象的机械理解。 （iii）识别和表征 允许高度突变细胞生存，增殖和进化的其他因素 特质。癌细胞必须维持巨大的突变负荷，因此有毒蛋白质组 不稳定。我将筛选蛋白质挽救高度突变菌株的生长，但不要 影响未分解的父母。 （iv）识别并表征其他基于蛋白质的 促进适应新环境的机制。使用数字核糖体 分析，酵母和细菌遗传学以及生物化学我将表征两个prions [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 这些研究将提供有关HSP90和蛋白质稳态的详细见解 通常，控制能够耐药性以及如何使用这些机制的信号通路 有助于突破性抵抗。他们还将暴露所有人共有的阿喀琉斯脚跟 癌症 - 对能够维持大规模突变负荷的因素上瘾。