Molecular Mechanism Governing Oxygen Signaling and Heme Regulation by Gis1

Gis1 控制氧信号传导和血红素调节的分子机制

基本信息

批准号：
9072488
负责人：
Li Zhang
金额：
$ 4.57万
依托单位：
UNIVERSITY OF TEXAS DALLAS
依托单位国家：
美国
项目类别：
财政年份：
2014
资助国家：
美国
起止时间：
2014-08-01 至 2019-02-28
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9072488
关键词：
Alzheimer&apos s Disease Amino Acids Anemia Cells Cessation of life Complex DNA DNA Binding DNA-Protein Interaction Data Disease Environment Eukaryota Event Goals Health Heme Histones Homologous Gene Human Hypoxia Hypoxia Inducible Factor In Vitro Injury Ischemia Life Liver Dysfunction Malignant Neoplasms Mediating Medical Membrane Proteins Mixed Function Oxygenases Molecular Mutagenesis Myocardial Infarction Nervous system structure Nuclear Organism Oxygen Pathogenesis Pathway interactions Porphyrias Proteins Public Health Radiation therapy Regulation Research Resistance development Science Signal Transduction Stroke Tertiary Protein Structure Testing Work Yeasts base chemotherapy design genetic regulatory protein improved innovation insight knowledge base member novel programs protein protein interaction research study response sensor tumor

项目摘要

DESCRIPTION (provided by applicant): There are fundamental gaps in understanding how regulatory proteins sense changes in oxygen and heme levels and promote cellular responses. Such gaps make it difficult to design effective strategies to treat diseases associated with hypoxia or heme dysregulation. The long- term goal is to better understand the molecular mechanisms by which the JmjC domain- containing proteins sense and promote cellular responses to changes in oxygen or heme levels. The objective of this application is to elucidate the molecular events underlying oxygen and heme regulation of Gis1 activity. The yeast Gis1 protein is a member of the JHDM3/JMJD2 subfamily of demethylases and a DNA-binding transcriptional regulator. Recent evidence shows that Gis1 is regulated by both oxygen and heme; Gis1 exemplifies a new class of regulators mediating heme and oxygen signaling. The central hypothesis is that the Gis1 JmjC domain cooperates with other domain(s) and proteins to sense and respond to changes in oxygen or heme levels and alter protein stability, protein-protein and protein-DNA interactions, thereby promoting oxygen and heme signaling. The rationale for the proposed work is that understanding how heme and oxygen regulate Gis1 can provide novel insights into how mammalian JmjC domain-containing proteins can be regulated by heme and oxygen and how their activity can be modulated to improve health. Guided by strong preliminary data, this central hypothesis will be tested by pursuing two specific aims: (1) Define the Gis1 modules mediating oxygen and heme signaling and characterize the direct interaction between Gis1 and heme; and (2) Characterize the effects of oxygen and heme on Gis1 protein interaction and DNA binding. Under the first aim, mutagenesis studies will be carried out to pinpoint Gis1 modules and residues critical for mediating oxygen and heme signaling in live cells, and in vitro studies will be carried out to examine the direct interaction between Gis1 and heme. Under the second aim, the effect of heme on Gis1 DNA binding will be examined, and purified Gis1 complexes will be characterized to determine the effect of oxygen and heme on Gis1 protein-protein interactions. The approach is innovative because it represents a new and substantive departure from the status quo in the studies of oxygen and heme signaling and of JmjC domain-containing demethylases. The proposed research is significant, because it is expected to advance the understanding of how the JmjC domain-containing proteins sense and respond to changes in oxygen and heme levels.

描述（由申请人提供）：了解调节蛋白如何感知氧气和血红素水平的变化并促进细胞反应的基本差距。这种差距使设计有效的策略很难治疗与缺氧或血红素失调相关的疾病。长期目标是更好地了解含有蛋白质的JMJC结构域感知的分子机制，并促进细胞对氧或血红素水平变化的反应。该应用的目的是阐明GIS1活性的氧气和血红素调节的分子事件。酵母GIS1蛋白是脱甲基酶的JHDM3/JMJD2亚家族和DNA结合转录调节剂的成员。最近的证据表明，GIS1受氧和血红素的调节。 GIS1例证了一类新的调节器，介导血红素和氧信号传导。中心假设是GIS1 JMJC结构域与其他结构域和蛋白质合作，以感知并应对氧或血红素水平的变化并改变蛋白质稳定性，蛋白质 - 蛋白质和蛋白-DNA相互作用，从而促进氧气和血红素信号传导。拟议工作的理由是，了解血红素和氧气如何调节GIS1如何提供新的见解，以了解如何通过血红素和氧气调节含有哺乳动物的JMJC结构蛋白，以及如何调节其活性以改善健康。在强大的初步数据的指导下，将通过追求两个具体目的来检验该中心假设：（1）定义介导氧和血红素信号传导的GIS1模块，并表征GIS1与Heme之间的直接相互作用；（2）表征氧和血红素对GIS1蛋白相互作用和DNA结合的影响。在第一个目的下，将进行诱变研究以查明GIS1模块和残基，对于介导活细胞中的氧和血红素信号至关重要，并将进行体外研究以检查直接相互作用在GIS1和Heme之间。在第二个目标下，将检查血红素对GIS1 DNA结合的影响，并将纯化的GIS1复合物进行表征，以确定氧和血红素对GIS1蛋白蛋白质相互作用的影响。这种方法具有创新性，因为它代表了氧气和血红素信号传导以及含JMJC结构域的脱甲基酶的新现状的新事物。拟议的研究很重要，因为有望提高人们对含JMJC结构域蛋白质的理解，并对氧气和血红素水平的变化做出反应。