LACTOGEN-REGULATED CALPACTIN GENE

催乳素（PRL）和相关激素在父母的照顾中起着重要作用。 后代和新陈代谢的调节。 在哺乳动物中，这包括 怀孕和哺乳的调节。 催乳素不仅控制 乳腺的催乳分化，而且还影响各种 其他靶组织。 其中许多影响对于支持 与妊娠相关的生长、分化和营养， 养育 这项研究计划的总体目标是了解分子 通过研究钙调素的刺激来研究PRL作用的生理学 在鸽子阴囊中的表达。 这种监管效果是一种富有成效的模式 系统中，同时检查两个分子机制， PRL诱导的基因表达和编码Ca ~（2+）的基因的功能 依赖性膜结合蛋白，它们是 癌基因和生长因子受体酪氨酸激酶。 我们将要研究的PRL调节基因（calpactin）本质上是 作为基础细胞生物学的一个组成部分， 研究PRL机制的有价值的终点。 这是一个积极的 该系统与PRL监管的其他系统（即， 乳蛋白基因），这些基因在哺乳动物中不起任何基本或一般作用， 细胞生理学 这些实验的目的集中在三个一般性主题上。 第一 我们对PRL诱导的钙调蛋白基因进行了全面的分析， 克隆的。 这些实验将确定独特的结构， 与这种形式的钙蛋白基因的诱导有关。 假设 要测试的将包括PRL诱导的calpactin基因被剪接， 或者它是从另一个启动子表达的 绝佳的价钱 将不受PRL调节的相关mRNA与 决定这个基因表达的特殊特征。 第二、 人钙蛋白的片段的重组，这些片段是 酪氨酸激酶与催乳素诱导的等同片段的调节 将制备和测试酪氨酸激酶调节的钙蛋白酶。 这些实验将检验生产性磷酸化 调节由下游的特定氨基酸序列决定， 人类钙调蛋白的磷酸化位点 第三，免疫定位 和中和calpactin mRNA表达 反义寡核苷酸将被用来检查这种功能， PRL诱导的增殖和分化中的蛋白质。

Prolactin (PRL) and related hormones play major roles in parental care of offspring and the regulation of metabolism. In mammals this includes regulation of pregnancy and lactation. Prolactin not only controls the lactogenic differentiation of mammary gland, but also affects a variety of other target tissues. Many of those effects are important to support the growth, differentiation, and nutrition associated with pregnancy and nurturing. The general goal of this research program is understanding the molecular physiology of PRL action by studying the stimulation of calpactin expression in pigeon cropsac. This regulatory effect is a productive model system in which to examine simultaneously both the molecular mechanisms of PRL-induced gene expression and the function of genes which encode Ca2+- dependent membrane-binding proteins that are major cellular substrates of oncogene and growth-factor receptor tyrosine kinases. The PRL-regulated gene which we will be examining (calpactin) is inherently interesting as a component of basic cellular biology, as well as being a valuable endpoint from which to study PRL mechanisms. This is one positive aspect which distinguishes this system from others regulated by PRL (i.e. milk protein genes) which do not play any fundamental or general roles in cellular physiology. The aims of these experiments are focussed on three general topics. First a comprehensive analysis of the PRL-induced calpactin gene which we have cloned. These experiments will determine the unique structures which are associated with induction of this form of the calpactin genes. Hypotheses to be tested will include that the PRL-induced calpactin gene is spliced in a specific manner or that it is expressed from an alternative promoter site. Related mRNAs which are not PRL-regulated will be compared to determine special features of this gene's expression. Second, recombinations of segments of a human calpactin which are sites of regulation by tyrosine kinases with equivalent segments of the PRL-induced calpactin which is to tyrosine kinase-regulated will be made and tested. These experiments will test the hypothesis that productive phosphorylative regulation is determined by specific amino acid sequences downstream from the phosphorylation site in human calpactin. Thirdly, immunolocalization of calpactin protein and neutralization of calpactin mRNA expression with antisense oligonucleotides will be used to examine the function of this protein in PRL-induced proliferation and differentiation.