MULTIHORMONAL REGULATION OF GENE EXPRESSION

这项提议的长期目标是理解复杂的 基因多激素调控中的调控事件 表达和细胞分化，特别强调 了解类固醇激素的作用机制。这个 鸡卵清蛋白基因的代表和优秀模式 研究是因为两种类固醇激素，雌激素和任何一秒 类固醇，与多肽激素胰岛素协同作用可增加 该基因在原代输卵管细胞中的表达超过200倍 文化。另一个蛋清基因的激素调节 溶菌酶将被用来区分基因特异性 监管机制和更普遍适用的事件。这个 本提案的具体目标是：(1)确定和描述 类固醇依赖的调控因素 鸡卵清蛋白和溶菌酶基因转录的初步研究 输卵管细胞，(II)研究多激素相互作用 调控卵清蛋白基因，定义和鉴定卵清蛋白基因。 卵清蛋白和溶菌酶基因中的调控元件 负责组织特异性表达，并(LV)确定如何 雌激素增加了靶基因的mRNAs的稳定性。 这些目标最初将依赖于基因转移技术来 确定卵清蛋白中的功能相关区域和 溶菌酶基因。基因已经被创造出来，包含了 卵清蛋白或溶菌酶基因的5‘侧翼区融合到 细菌氯霉素乙酰转移酶的标记基因 (CAT)。删除和连接子扫描突变体将在 融合基因，并将它们导入初级输卵管 手机。细胞将在适当的荷尔蒙环境中培养。 在检测转基因和内源性基因的表达之前 蛋清基因。一旦确定了监管要素， 研究将集中在分离和鉴定能够 绑定到这些区域。 关于类固醇激素作用机制的信息 而关于基因表达的多激素调节应该 事实证明，这对于了解复杂的 高等真核生物。类固醇激素和生长因子 与生殖道癌的病因学有关 乳腺和前列腺等组织，更好地了解 这些分子事件只会改善治疗和 这些恶性肿瘤的预防。另外，这样的知识 可能会为更好地开发 避孕药具和治疗生殖障碍的援助。

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.