RUI: DNA-Protein Interactions Regulating Transcription of the Skeletal Muscle Actin Gene

RUI：DNA-蛋白质相互作用调节骨骼肌肌动蛋白基因的转录

• 批准号: 9604289
• 负责人: Sandra Sharp
• 金额: $ 27万
• 依托单位: California State L A University Auxiliary Services Inc.
• 依托单位国家: 美国
• 项目类别: Continuing Grant
• 财政年份: 1997
• 资助国家: 美国
• 起止时间: 1997-02-15 至 2000-01-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=9604289&HistoricalAwards=false
• 关键词: RUI; DNA; Protein; Interactions; Regulating

9604289 Sharp Two families muscle-specific transcription factors - the myogenesis regulatory factor (MRF) family which is expressed only in skeletal muscle, and the myocyte enhancer factor-2 (MEF2) family which is expressed in all muscle types - are required for skeletal myogenesis in mammals, and have been shown to act synergistically. Members of each family bind to specific DNA sequences in the promoter regions of muscle-specific genes and activate transcription. This research will determine how these two families of transcription factors act synergistically to activate the transcription of the skeletal muscle (skm) actin gene. Unlike other muscle-specific genes which demonstrate MRF/MEF2 synergism, the MRF and MEF2 binding sites of the skm actin gene are physically separated from each other on the DNA. Co-transfection and mutagenic analysis will be performed using an skm actin gene promoter-chloramphenicol acetyltransferase reporter gene construct. Co-transfection into non-muscle cells of this reporter gene with DNA constructs expressing specific MRF and/or MEF2 transcription factors will be used to evaluate the effect on transcription of particular combinations of MRF and MEF2 proteins. The role of specific DNA binding sites within the skm actin gene promoter will be determined using the same reporter gene in which specific MRF or MEF2 binding sites have been mutated. The results of this research will provide the basis for future analysis of the promoter-specific and transactivator-specific regions of the MRF and MEF2 proteins, and should lead both to broader regulatory models that can be tested in the context of other promoters and to a better understanding of the process of myogenesis. The process by which skeletal muscle precursor cells develop into skeletal muscle is regulated by an intricate interplay of molecular mechanisms. One approach to understanding these mechanisms is to study the DNA-protein interactions which regulate the expression of muscle-specific genes during development. This research will determine how two families of muscle-specific proteins which are required for skeletal muscle development act synergistically to activate the expression of the skeletal muscle actin gene. The results of this research should lead both to broader regulatory models that can be tested in the context of other genes and to a better understanding of the process of skeletal muscle development.

两个家族的肌肉特异性转录因子-仅在骨骼肌中表达的肌生成调节因子（MRF）家族和在所有肌肉类型中表达的肌细胞增强因子-2（MEF 2）家族-是哺乳动物骨骼肌生成所需的，并且已经显示出协同作用。 每个家族的成员与肌肉特异性基因的启动子区域中的特异性DNA序列结合并激活转录。 这项研究将确定这两个家族的转录因子如何协同作用，以激活骨骼肌（skm）肌动蛋白基因的转录。 与其他显示MRF/MEF 2协同作用的肌肉特异性基因不同，skm肌动蛋白基因的MRF和MEF 2结合位点在DNA上彼此物理分离。 将使用skm肌动蛋白基因启动子-氯霉素乙酰转移酶报告基因构建体进行共转染和致突变分析。 将该报告基因与表达特异性MRF和/或MEF 2转录因子的DNA构建体共转染到非肌肉细胞中，将用于评价MRF和MEF 2蛋白的特定组合对转录的影响。 skm肌动蛋白基因启动子内特异性DNA结合位点的作用将使用其中特异性MRF或MEF 2结合位点已经突变的相同报告基因来确定。 这项研究的结果将为未来分析MRF和MEF 2蛋白的启动子特异性和反式激活子特异性区域提供基础，并应导致更广泛的监管模式，可以在其他启动子的背景下进行测试，并更好地了解肌生成的过程。 骨骼肌前体细胞发育成骨骼肌的过程受到复杂的分子机制相互作用的调节。 理解这些机制的一种方法是研究在发育过程中调节肌肉特异性基因表达的DNA-蛋白质相互作用。 这项研究将确定骨骼肌发育所需的两个肌肉特异性蛋白家族如何协同激活骨骼肌肌动蛋白基因的表达。这项研究的结果应该导致更广泛的监管模式，可以在其他基因的背景下进行测试，并更好地了解骨骼肌发育的过程。