MULTIHORMONAL REGULATION OF GENE EXPRESSION

本提案的长期目标是了解复杂的 基因的多激素控制中的调节事件 表达和细胞分化，特别强调 了解类固醇激素的作用机制。 的 鸡卵清蛋白基因代表和优秀的模型，这些 因为两种类固醇激素，雌激素和任何第二种 类固醇，协同肽激素胰岛素增加 该基因在原代输卵管细胞中表达超过200倍 cultures. 另一个卵白色基因的激素调节 溶菌酶将用于区分基因特异性 监管机制和更普遍适用的事件。 的 本提案的具体目标是：（1）确定和描述 负责类固醇依赖性的调节元件 卵清蛋白和溶菌酶基因的转录 输卵管细胞，（II）研究多激素相互作用， 调节卵清蛋白基因，（III）定义和表征 卵清蛋白和溶菌酶基因中的调节元件 负责组织特异性表达，和（IV）确定如何 雌激素增加靶基因mRNA的稳定性。 这些目标最初将依赖于基因转移技术， 鉴定卵清蛋白中的功能相关区域， 溶菌酶基因 基因已经被创造出来， 卵清蛋白或溶菌酶基因的5 '侧翼区融合到 标记基因细菌氯霉素乙酰转移酶 （CAT）。 缺失和接头扫描突变体将在 融合基因，并将它们转染到原代输卵管中 cell. 细胞将在适当的激素环境中培养 在测定转染的和内源的 卵白色基因。 一旦确定了监管要素， 研究将集中在分离和表征蛋白质， 绑定到这些区域。 关于类固醇激素作用机制的信息 关于基因表达的多激素调节， 对于理解复杂的监管事件， 高等真核生物 类固醇激素和生长因子 与生殖系统癌症的病因有关， 乳腺和前列腺等组织，更好地了解 这些分子事件只能改善治疗， 预防这些恶性肿瘤。 此外，这些知识 可以提供更好的发展洞察力 避孕药和辅助治疗生殖疾病。

The long range goal of this proposal is to understand the complex regulatory events involved in the multihormonal control of gene expression and cellular differentiation with particular emphasis on understanding the mechanism of action of steroid hormones. The chicken ovalbumin gene represents and excellent model for these studies because two steroid hormones, estrogen and any second steroid, synergize with the peptide hormone insulin to increase expression of this gene over 200-fold in primary oviduct cell cultures. The hormonal regulation of another egg white gene lysozyme will be used to distinquish between gene-specific regulatory mechanisms and more generally applicable events. The specific aims of this proposal are to (1) identify and characterize the regulatory elements responsible for steroid-dependent transcription of the ovalbumin and lysozyme genes in primary oviduct cells, (ll) investigate the multihormonal interactions tht regulate the ovalbumin gene, (lll) define and characterize the regulatory elements in the ovalbumin and lysozyme genes responsible for tissue -specific expression, and (lV) determine how estrogen increases the stability of mRNAs from target genes. These objectives will rely initially on gene trasfer techniques to identify functionally relevent regions in the ovalbumin and lysozyme genes. Genes have already been created containing the 5'-flanking regions of the ovalbumin or lysozyme genes fused to the marker gene bacterial chloramphenicol acetyltransferase (CAT). Deletion and linker scanning mutants will be made in the fusion genes, and they will be transfected into primary oviduct cell. The cells will be cultured in the appropriate hormonal milieu prior to assay for expression of the transfected and endogenous egg white genes. Once regulatory elements have been identified, studies will focus on isolating and characterizing the proteins that bind to these regions. Information about the mechanism of action of steroid hormones and about the multihormonal regulation of gene expression should prove invaluable for understanding complex regulatory events in higher eucaryotes. As steroid hormones and growth factors have been implicated in the etiology of cancers of the reproductive tissues such as breast and prostate, a better understanding of these molecular events can only improve the treatment and prevention of these malignancies. In addition, such knowledge may provide insights into the development of better contraceptives and aid in the treatment of reproductive disorders.