Unusual mechanisms of metal regulation down to single-cell single-molecule level

在此输入文本，它是应用程序的新摘要信息。此部分不得超过30行文本。 确定细胞如何调节锌和铜等过渡金属的吸收和流出是阐明金属稳态细胞机制的关键组成部分。细菌模型系统为理解金属响应基因调控提供了范例。在大肠在大肠杆菌中，金属调节剂ZntR感知Zn过量并激活Zn流出，而Zur感知Zn充足并抑制Zn吸收，以将这种必需金属保持在细胞中适当的生理水平。CueR是ZntR的同源物，感应细胞内的Cu以激活Cu流出/解毒基因，以保持这种有毒金属最小。这里的长期目标是了解细胞中的金属调节如何用于预防和治疗目的。为了实现这一目标，PI已经建立了一个国际上独特的研究计划，该计划应用和开发先进的单分子单细胞方法来询问和了解体外和活细胞中细菌金属调节的机制，通过大量生物化学/生物物理学和蛋白质/基因工程方法以及与生物学家建立的合作进一步增强。这项研究发现了Cu/Zn响应转录调控的第一种机制，但也出现了新的问题。此次更新的目的是继续研究细菌中金属调节的机制。这项研究的前提包括（细菌）金属调节在生物学中的重要性，发现的新的和广泛相关的调节机制，以及结合单分子/细胞和批量测量的力量。本研究包括几个具体的目标，每个目标都有子目标，即研究大肠杆菌中锌敏感金属调节蛋白Zur和铜敏感双组分系统CusRS的作用机制。大肠杆菌，以及细胞与细胞之间的相互作用对单个细胞的Zu调节。这项研究意义重大，因为它将阐明金属调节剂在调节金属流出和吸收方面的新分子机制，并提供有关金属细胞生物学的基本知识，用于确定涉及类似调节过程的疾病的原因或开发预防措施，并帮助开发（生物）化学策略来操纵细菌Zn/Cu调节以损害病原体生长。这项研究是创新的，因为它应用/开发了新的单分子操作，成像和分析方法，并在转录调控中引入了新的机制概念。

Enter the text here that is the new abstract information for your application. This section must be no longer than 30 lines of text. Defining how cells regulate the uptake and efflux of transition metals such as Zn and Cu is a key component in elucidating cellular mechanisms of metal homeostasis. Bacterial model systems provide paradigms for understanding metal-responsive gene regulation. In E. coli, the metalloregulator ZntR senses Zn excess and activates Zn efflux, while Zur senses Zn sufficiency and represses Zn uptake, to keep this essential metal at appropriate physiological levels in the cell. CueR, a homolog of ZntR, senses intracellular Cu to activate Cu efflux/detoxification genes to keep this toxic metal minimal. The long-term goal here is to understand how metal regulation in the cell can be manipulated for preventive and therapeutic purposes. Toward this goal, the PI has established an internationally unique research program that applies and develops advanced single- molecule single-cell approaches to interrogate and understand the mechanisms of bacterial metal regulation both in vitro and in live cells, which are further enhanced by bulk biochemical/biophysical and protein/genetic engineering approaches and established collaborations with biologists. The research has led to discoveries of first-of-their-kind mechanisms of Cu/Zn-responsive transcriptional regulation, but new questions also emerged. The objective of this renewal is to continue this program in studying the mechanism of metal regulation in bacteria. The premise of this research comprises the importance of (bacterial) metal regulation in biology, the discovered novel and broadly relevant regulation mechanisms, and the power of combining single- molecule/cell and bulk measurements. The proposed research contains a few specific aims, each with sub- aims, to study the mechanism of Zn-sensing metelloregulator Zur and the Cu-sensing two component system CusRS in E. coli, as well as the cell-cell interactions on the Zu regulations of individual cells. The research is significant because it will elucidate novel molecular mechanisms of metalloregulators in regulating metal efflux and uptake, as well as provide fundamental knowledge about cell biology of metals in general, for identifying causes or developing preventions of diseases that involve similar regulation processes, and for helping the development of (bio)chemical strategies to manipulate bacterial Zn/Cu regulation to impair pathogen growth. The research is innovative because it applies/develops novel single-molecule manipulation, imaging, and analysis methods, and introduce new mechanistic concepts in transcription regulation.