OXYGEN SENSING BY THE RESPIRATORY CHAIN

通过呼吸链进行氧气感应

• 批准号: 6636339
• 负责人: Kurt Eric Kwast
• 金额: $ 21.94万
• 依托单位: UNIVERSITY OF ILLINOIS AT URBANA-CHAMPAIGN
• 依托单位国家: 美国
• 财政年份: 2001
• 资助国家: 美国
• 起止时间: 2001-04-01 至 2006-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6636339
• 关键词: DNA binding protein; Saccharomyces cerevisiae; beta galactosidase; biological signal transduction; carbon monoxide; cellular respiration; chemical structure function; cytochrome oxidase; functional /structural genomics; fungal genetics; fungal proteins; gel electrophoresis; gene environment interaction; genetic regulatory element; hemoprotein; intermolecular interaction; mitochondria; molecular cloning; molecular site; northern blottings; nucleic acid sequence; oxygen; polymerase chain reaction; yeast two hybrid system

DESCRIPTION (Applicant's abstract): One of the challenges facing researchers working on oxygen-regulated biological phenomenon, as well as medical practitioners attempting to minimize hypoxia-induced tissue damage, is gaining a fundamental understanding of the molecular mechanisms for gauging oxygen availability. In spite of significant progress in our understanding of the physiological response to hypoxia, the molecular components involved in sensing oxygen and the pathways for transducing this information are poorly understood. The long-term objective of our research is to delineate oxygen-sensing mechanisms and the signaling pathways involved in eliciting adaptive responses at the level of gene expression. In this revised proposal, we focus on determining the role that cytochrome c oxidase, and possibly other components of the mitochondrial respiratory chain, plays in oxygen-sensing pathways. A yeast model will be used to address the following specific aims. Aim 1: Identify the cis site(s) in the promoters of oxygen-responsive genes that confer(s) sensitivity to inhibitors and mutations in the respiratory chain. To address this aim, the promoter regions of several oxygen-responsive genes will be fused to the reporter gene LacZ and used to define the minimal-sized DNA fragment and essential nucleotides that confer sensitivity to inhibitors and mutations in the respiratory chain. Aim 2: Identify trans-acting factors that interact with these cis sites and other proteins involved in transducing the low oxygen signal from the mitochondrion to the nucleus. Once the cis site is defined, one-hybrid and two-hybrid screens will be used to identify the trans-acting factor(s) and other proteins responsible for mediating sensitivity to inhibitors and mutations in the respiratory chain. In addition, genome-wide transcript profiling analyses will be used to identify additional sensory and signaling proteins. These studies can be expected to provide novel findings and new insight into the crosstalk between the mitochondrion and nucleus that is involved in modulating the expression of oxygen-responsive genes and provide a general model of oxygen sensing in eukaryotic cells.

描述（申请人的摘要）：研究人员面临的挑战之一 从事氧气调节的生物学现象以及医学 试图最大程度减少缺氧引起的组织损伤的从业者正在获得 对测量氧气的分子机制的基本理解 可用性。尽管我们对 对缺氧的生理反应，参与感应的分子成分 氧气和传递此信息的途径知之甚少。 我们研究的长期目的是描述氧气感应 引起自适应反应的机制和信号通路 在基因表达水平。在这项修订的建议中，我们专注于 确定细胞色素C氧化酶以及可能其他成分的作用 线粒体呼吸链的作用，在氧气感应途径中发挥作用。一个 酵母模型将用于解决以下特定目标。 目标1：识别氧响应基因启动子中的顺式位点 这种赋予抑制剂和突变的敏感性 链。 为了解决这个目标，几个氧气响应基因的启动子区域 将融合到记者基因lacz，并用于定义最小尺寸 DNA片段和必需的核苷酸，赋予抑制剂敏感性 和呼吸链中的突变。 目标2：确定与这些顺式位点相互作用的反式作用因素和 其他参与传递低氧信号的蛋白质 线粒体向细胞核。 一旦定义了顺式站点，将使用一杂交和两杂交屏幕 确定跨作用因子和其他负责的蛋白质 介导呼吸链中抑制剂和突变的敏感性。在 此外，全基因组的成绩单分析分析将用于识别 其他感觉和信号蛋白。 可以期望这些研究提供新颖的发现和新的见解 线粒体和核之间的串扰 调节氧响应基因的表达并提供一般 真核细胞中的氧气传感模型。