MECHANISMS AND MACROMOLECULAR INTERACTIONS UNDERLYING CELLULAR RESPONSES TO STRESS SIGNALS

细胞对应激信号反应的机制和大分子相互作用

• 批准号: 10570860
• 负责人: Alexandra M. Deaconescu
• 金额: $ 39.88万
• 依托单位: BROWN UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-04-01 至 2027-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10570860
• 关键词: Address; Affect; Age; Antibiotics; Antimicrobial Resistance; Area; Bacteria sigma factor KatF protein; Biology; COVID-19 pandemic; Catalysis; Collaborations; Collection; Combination Drug Therapy; Coupled; Coupling; DNA; DNA Damage; DNA Repair; DNA-Directed RNA Polymerase; Development; Dissection; Environment; Evolution; Genetic Transcription; Genomics; Infection; Lesion; Mediating; Modality; Molecular Evolution; Mutagenesis; Nature; Nucleotide Excision Repair; Organism; Pathway interactions; Process; Proteins; Proteolysis; Public Health; Reagent; Regulation; Research; SOS Response; Signal Transduction; Specificity; Stress; Structural Biochemistry; Transcription Elongation; Transcription Initiation; Transcription-Coupled Repair; Work; antimicrobial; biological adaptation to stress; cross immunity; novel; pathogen; programs; promoter; recruit; repaired; response; stressor; structural biology; transmission process

PROJECT SUMMARY Proteins carrying out DNA repair, replication and transcription, processes essential for the stable transmission and expression of genomic information, share the same track – the DNA – and require delicate coordination to achieve their function. Often, regulation of these processes occurs at the level of targeting or well-timed recruitment, but also catalysis and even turnover of the factor in question. We will address these three principal modalities of regulation in the context of bacterial responses to stress signals. Our questions and hypotheses are grouped into two large areas, (1) regulation of the general stress response mediated by the promoter specificity subunit RpoS; this is exemplary of a general stress response mechanism that affect transcription initiation, and (2) specific responses to DNA damage such as subpathways of nucleotide excision repair; these are exemplary of how the DNA damage response interfaces with transcription elongation, termination as well as replication. The principal questions that will be addressed by my research plan are: A. How are activating and inhibitory regulatory inputs integrated to tune the RpoS core pathway and globally reprogram transcription in response to an adverse environment? How does the ClpXP adaptor RssB and stress-specific ClpXP anti-adaptors tune the proteolysis of RpoS in a stress-specific manner? B. What are the mechanisms for preferential recruitment of the NER machinery to specific lesions, or to the template strand, directly read by RNA polymerase? How is the availability of early NER factors regulated by proteolysis in the context of DNA damage and transcription-coupled DNA repair? How and when do transcription-repair coupling factors collaborate with key players in the SOS response to promote mutagenesis? Can we leverage these mechanisms towards developing novel anti-evolution drugs for combination therapies against infection with diverse pathogens? Previous work has already allowed us to build a critical collection of reagents and expertise in the structural biology and biochemistry of transcription-coupled repair and RpoS biology. This gives us now an excellent entry point for a mechanistic dissection of the processes listed above and the development of novel antimicrobial strategies. The significance of our work is thus not only fundamental and conceptual in nature, but has immediate applications in controlling the worldwide public health crisis of antimicrobial resistance, particularly in the age of the COVID-19 pandemic.

项目总结 进行DNA修复、复制和转录的蛋白质，是稳定传播所必需的过程 和表达基因组信息，共享相同的轨迹-DNA-并且需要微妙的协调来 实现它们的功能。通常，对这些过程的监管是在定向或适时的水平上进行的 招聘，也是催化甚至换人的问题因素。我们将解决这三个原则 细菌对胁迫信号反应的调控方式。我们的问题和假设 可分为两大块：(1)启动子介导的一般应激反应的调节 特异性亚基rpos；这是影响转录的一般应激反应机制的范例。 以及(2)对DNA损伤的特异性反应，如核苷酸切除修复的亚途径；这些 是DNA损伤反应如何与转录延伸、终止以及 复制。我的研究计划将解决的主要问题是： A.如何整合激活和抑制调节输入来调节rpos核心通路和 在全球范围内重新编程转录以应对不利环境？ClpXP如何 适配器RSSB和应激特异的ClpXP反适配器调节应激特异的RPO的蛋白质分解 举止？ B.将NER机制优先招募到特定皮损的机制是什么？ 或者直接被RNA聚合酶读取的模板链？早期NER的可用性如何 DNA损伤和转录偶联DNA修复中的蛋白降解调节因子？ 转录修复偶联因子如何以及何时与SOS反应中的关键角色协作 来促进基因突变吗？我们能否利用这些机制来开发新的反进化技术 针对不同病原体感染的联合治疗药物？ 以前的工作已经使我们能够在结构上建立一个关键的试剂和专业知识集合 转录偶联修复和rpos生物学的生物学和生物化学。这给了我们一个很好的入口 从机理上剖析上述过程和开发新型抗菌剂的要点 战略。因此，我们工作的意义不仅是根本性的和概念性的，而且具有直接的 在控制抗菌素耐药性的全球公共卫生危机方面的应用，特别是在 新冠肺炎大流行。