MEDIUM CHAIN ACYL-COA DEHYDROGENASE GENE EXPRESSION

中链酰基辅酶A脱氢酶基因表达

基本信息

批准号：
2144651
负责人：
DANIEL PATRICK KELLY
金额：
$ 11.25万
依托单位：
WASHINGTON UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
1992
资助国家：
美国
起止时间：
1992-09-30 至 1996-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/2144651
关键词：
DNA binding protein acyl coA dehydrogenases chloramphenicol acetyltransferase fatty acid metabolism flavoproteins gene expression genetic promoter element genetic regulatory element genetically modified animals human tissue laboratory mouse laboratory rat plasmids retinoate retinoid binding proteins tissue /cell culture transcription factor transfection

项目摘要

Medium-chain acyl-CoA dehydrogenase (MCAD,EC 1.3.99.3) is a mitochondrial flavoenzyme which catalyzes the initial, rate-limiting, reaction in fatty acid beta-oxidation. Inherited MCAD deficiency, a cause of fasting coma, liver dysfunction, and sudden death in childhood, reflects the importance of this enzyme in energy metabolism. The MCAD gene is highly regulated, in parallel with fatty acid oxidation rates, among tissues and during development. Elucidation of the mechanisms involved in the regulation of the MCAD gene will provide insights relevant to MCAD deficiency and to the understanding of mechanisms involved in the regulation of expression of nuclear genes encoding mitochondrial enzymes involved in fatty acid metabolism under normal conditions and in a variety of physiologic and disease states. The major goals of this proposal include the structural and functional characterization of the promoter and upstream cis-acting regulatory regions of the human MCAD gene including identification of the elements involved in tissue-specific, developmental, and retinoic acid-responsive transcriptional regulation. Ultimately, we hope to identify regulatory sequences and transacting regulatory DNA binding proteins involved in coordinate control of genes encoding metabolic enzymes and mitochondrial proteins. The promoter region and regulatory elements will be characterized by transfecting a variety of chimeric plasmids containing varying lengths of MCAD gene 5'-flanking DNA fused to the bacterial chloramphenicol acetyltransferase (CAT) gene into mammalian cells derived from several tissues with differing MCAD mRNA expression levels. Retinoic acid response elements will be localized by identifying the regulatory sequences of the MCAD gene which confer retinoic acid-responsive transcriptional activation to the MCAD-CAT plasmids in mammalian cells in culture. The regulatory sequences will be compared to sequences of known regulatory elements in other genes, particularly those encoding enzymes involved in metabolism. The regulatory elements will be further characterized by performing DNA-nuclear protein binding assays. Ultimately, the trans-acting regulatory proteins involved in the transcriptional regulation of the MCAD gene will be identified by isolating and characterizing their cDNAs. The role of the regulatory elements in developmental and tissue-specific regulation of the MCAD gene in vivo will be evaluated by employing a transgenic mouse system.

中链酰基-COA脱氢酶（MCAD，EC 1.3.99.3）是线粒体黄油酶在脂肪中催化初始速率，限速性反应酸β-氧化。继承的MCAD缺乏症，禁食的原因，肝功能障碍和童年突然死亡反映了重要性能量代谢中的这种酶。 MCAD基因受到高度调节，与脂肪酸氧化速率以及组织之间和期间发展。阐明与调节有关的机制 MCAD基因将提供与MCAD缺乏症相关的见解和对表达调节涉及的机制的理解核基因编码参与脂肪酸的线粒体酶在正常条件下以及各种生理和疾病状态。该提案的主要目标包括结构性以及启动子和上游顺式作用的功能表征人MCAD基因的调节区域包括鉴定涉及组织特异性，发育和视网膜的元素酸反应性转录调控。最终，我们希望确定调节序列和交易调节DNA结合参与编码代谢的基因的坐标控制的蛋白质酶和线粒体蛋白。启动子区域和监管元素的特征是转染各种嵌合含有不同长度的MCAD基因5'Fanking DNA的质粒融合细菌氯霉素乙酰基转移酶（CAT）基因进入哺乳动物源自具有不同MCAD mRNA表达的几个组织的细胞水平。视黄酸反应元件将通过识别来定位 MCAD基因的调节序列，赋予视网膜对MCAD-CAT质粒的酸反应转录激活培养中的哺乳动物细胞。调节序列将与其他基因中已知调节元件的序列，尤其是那些基因的序列编码与代谢有关的酶。监管元素将是进一步以执行DNA-核蛋白结合测定的特征。最终，与 MCAD基因的转录调节将通过隔离和表征其cDNA。监管的作用 MCAD基因的发育和组织特异性调节元素体内将通过使用转基因小鼠系统来评估。

项目成果

期刊论文数量（21）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Total skeletal muscle PGC-1 deficiency uncouples mitochondrial derangements from fiber type determination and insulin sensitivity.

DOI：
10.1016/j.cmet.2010.11.008
发表时间：
2010-12-01
期刊：
Cell metabolism
影响因子：
29
作者：
Zechner C;Lai L;Zechner JF;Geng T;Yan Z;Rumsey JW;Collia D;Chen Z;Wozniak DF;Leone TC;Kelly DP
通讯作者：
Kelly DP

A pleiotropic element in the medium-chain acyl coenzyme A dehydrogenase gene promoter mediates transcriptional regulation by multiple nuclear receptor transcription factors and defines novel receptor-DNA binding motifs.

中链酰基辅酶 A 脱氢酶基因启动子中的多效性元件介导多个核受体转录因子的转录调节，并定义了新的受体-DNA 结合基序。