Structural and functional basis of bacterial transcriptional regulation

细菌转录调控的结构和功能基础

• 批准号: 10712023
• 负责人: Oriana S Fisher
• 金额: $ 36.96万
• 依托单位: LEHIGH UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-09-01 至 2028-07-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10712023
• 关键词: Antibiotic Resistance; Antibiotics; Bacillus subtilis; Bacteria; Binding; Biochemical; Biochemical Process; Biochemistry; Biological Models; Copper; Development; Environment; Enzymes; Exhibits; Family; Family member; Future; Goals; Investigation; Kinetics; Microbiology; Molecular; Organism; Pathogenicity; Pathway interactions; Phosphotransferases; Phylogeny; Physiological; Process; Proteins; Public Health; Research; Specificity; Substrate Specificity; Transcription Repressor; Transcriptional Regulation; Transition Elements; Variant; biological adaptation to stress; computer studies; design; drug discovery; novel; novel therapeutics; preference; small molecule inhibitor; structural biology; tool; uptake

Project Summary / Abstract Antibiotic resistance poses an increasingly prevalent public health challenge, but advances in the development of new drugs has not progressed commensurately. A significant impediment lies in that many fundamental biochemical processes in bacteria remain poorly understood. The primary goal of the proposed research is to illuminate how bacteria respond and adapt to changes within their environments from a structural and mechanistic standpoint. Using the genetically tractable Gram positive organism Bacillus subtilis as our model system, we will focus our efforts on elucidating the molecular basis for copper-dependent transcriptional regulation and uncovering the drivers of enzyme specificity during the environmental stress response. Copper is required for bacterial survival, but excess levels of this transition metal can be toxic. Therefore, copper levels must be carefully controlled. The mechanisms by which copper export occurs have been extensively studied, but relatively little is known about copper import. We hypothesize that Cu uptake is regulated by Cu-dependent transcriptional repressors and the proteins under their control. Here, we will focus on a suite of proteins implicated in these processes, studying them at a detailed structural, molecular, and cellular level to better understand how they regulate copper uptake. In parallel, we will investigate what causes a family of very closely related kinases to specifically regulate distinct functions. Many bacterial species, including Bacillus subtilis and a number of pathogenic strains, encode one or more kinases that contain a domain termed the Bergerat fold. Despite their structural commonalities, enzymes with this domain exhibit strong preferences for their physiological binding partners. However, the molecular basis for how such specificity is conferred has yet to be investigated. We hypothesize that variations within the Bergerat fold influence substrate specificity and kinetics and can be targeted by small molecule inhibitors. By integrating tools from structural biology, microbiology, biochemistry, and computational studies, we will be poised to reveal new paradigms in transcriptional control. The completion of the proposed studies will catalyze our future investigations into elucidating analogous pathways in pathogenic strains, uncovering novel members of these families in bacterial phylogeny, and designing small molecule inhibitors.

项目总结/摘要 抗生素耐药性构成了日益普遍的公共卫生挑战，但 新药的开发进展缓慢。一个重要的障碍在于， 细菌的基本生化过程仍然知之甚少。建议的主要目标 研究的目的是阐明细菌如何从结构上适应环境的变化， 机械的观点。使用遗传上易处理的革兰氏阳性菌枯草芽孢杆菌作为我们的模型 系统，我们将集中精力阐明铜依赖性转录的分子基础 调节和揭示酶特异性的驱动程序在环境应激反应。铜是 细菌生存所必需的，但过量的这种过渡金属可能是有毒的。因此，铜含量 必须小心控制。铜输出发生的机制已被广泛研究， 但对铜的进口知之甚少。我们推测，铜的吸收是由铜依赖的 转录抑制因子及其控制下的蛋白质。在这里，我们将重点放在一套蛋白质牵连 在这些过程中，在详细的结构，分子和细胞水平上研究它们，以更好地了解它们是如何 它们调节铜的吸收。与此同时，我们将研究是什么原因导致一个非常密切相关的激酶家族 来具体调节不同的功能。许多细菌物种，包括枯草芽孢杆菌和一些 致病性菌株编码一种或多种激酶，所述激酶含有称为Bergerat折叠的结构域。尽管他们 结构上的共性，具有该结构域的酶表现出强烈的生理结合偏好 伙伴然而，如何赋予这种特异性的分子基础还有待研究。我们 假设Bergerat折叠内变化影响底物特异性和动力学， 被小分子抑制剂靶向。通过整合结构生物学，微生物学，生物化学， 和计算研究，我们将准备揭示转录控制的新范式。完成 的拟议研究将促进我们未来的调查，阐明类似的途径，在致病性 菌株，发现这些家族在细菌遗传学中的新成员，设计小分子 抑制剂的