HORMONAL AND CAMP REGULATION OF PROTEIN SYNTHESIS

多肽激素对蛋白质的生物合成起快速、特异的作用 以及目标组织中的基因表达，这对正常 发展和动态平衡。定义精确的生化机制 负责这一规定的人将提供一个概念性的框架， 以此来研究多种疾病的失调规律。促甲状腺激素 促肾上腺皮质激素释放激素、表皮生长因子、蛙皮素、佛波醇 酯(TPA)和cAMP在加入克隆大鼠后几分钟内起作用 垂体细胞培养(GH4)以产生特异性转录 催乳素基因的刺激；生长激素释放因子发挥作用 对生长激素基因有类似的作用。大鼠催乳素的调节部位 因此，生长激素(GH)的生物合成是由多肽激素 在基因转录水平上。我们建议定义精确的 基因组序列中不到200个碱基的5‘侧翼信息 定量转移多肽与类固醇激素调节 催乳素和生长激素基因的组合调控特性 包括DNA介导的融合基因转移的分析 转基因催乳素和生长激素基因组片段转化为 同源细胞系和原代培养的垂体细胞，并进入 使用适当的逆转录病毒载体系统的正常组织。几个 将使用方法来定义传授的序列 组织特异性神经内分泌基因的表达及鉴定关键 反式作用因素。亲和层析及一种新的表达 将采用策略来隔离互动式调控因素 对荷尔蒙和发育调节的基因模式至关重要 并克隆其编码的mRNAs。转基因动物 将使用各种方法来进一步定义序列的复杂模式 识别和染色体位置效应决定了 组织特异性发育模式的基因表达特征 神经内分泌系统。两种基因的结构和调控功能 快速磷酸化的新碱性染色质相关蛋白 作为对多肽激素的反应，cAMPL将通过 重组DNA技术。基于EGF受体的克隆 本实验室的cDNAs和多态RNA产物的鉴定， 我们建议对EGF的结构成分进行遗传学评估 生长、蛋白质合成和转化所需的受体。

Polypeptide hormones exert rapid, specific effects on protein biosynthesis and gene expression in their target tissues, critical for normal development and homeostasis. Defining the precise biochemical mechanisms responsible for this regulation would provide a conceptual framework with which to investigate many diseases of disorders regulation. Thyrotropin releasing hormone (TRH), epidermal growth factor (EGF), bombesin, phorbol esters (TPA), and cAMP act within minutes of their addition to clonal rat pituitary cell cultures (GH4) to produce specific transcriptional stimulation of the prolactin gene; growth hormone releasing factor exerts comparable actions on the GH gene. The site of regulation of rat prolactin and growth hormone (GH) biosynthesis by polypeptide hormones is therefore at the level of gene transcription. We propose to define the precise genomic sequences in the less than 200 bp of 5' flanking information which quantitatively transfer polypeptide and steroid hormonal regulation characteristic of prolatin and GH gene regulation using a combinatorial analysis including DNA-mediated gene transfer of fusion genes containing genetically modified prolactin and growth hormone genomic fragments into homologous cell lines and into primary pituitary cell cultures, and into normal tissues using appropriate retroviral vector systems. Several approaches will be utilized to define the sequences imparting tissue-specific neuroendocrine gene expression and to identify the critical trans-acting factors. Affinity chromatography and a novel expression strategy will be adapted to isolate the transactive regulatory factors critical for hormonally- and developmentally- mediated patterns of gene expression, and to clone their encoding mRNAs. Transgenic animal approaches will be used to further define the complex patterns of sequence recognition and chromosomal position effects which adjudicate the tissue-specific developmental patterns of gene expression characteristic of the neuroendocrine system. The structure and regulatory function of two novel basic chromatin associated proteins which are rapidly phosphorylated in response to polypeptide hormones aned cAMPL will be elucidated via recombinant DNA technology. Based upon the cloning of the EGF receptor cDNA in this laboratory and the identification of polymorphic RNA products, we propose genetic evaluation of the structural components of the EGF receptor required for growth, protein synthesis, and transformation.