GATA-BINDING PROTEINS REGULATE TESTICULAR GENE EXPRESSION

GATA 结合蛋白调节睾丸基因表达

• 批准号: 6240909
• 负责人: CHING-LING C CHEN
• 金额: $ 22.4万
• 依托单位: POPULATION COUNCIL
• 依托单位国家: 美国
• 财政年份: 1997
• 资助国家: 美国
• 起止时间: 1997-04-07 至 1998-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6240909
• 关键词: DNA footprinting; Leydig cells; Sertoli cells; binding proteins; gel mobility shift assay; gene expression; genetic promoter element; genetic regulation; hormone regulation /control mechanism; immunocytochemistry; in situ hybridization; inhibin; laboratory rabbit; laboratory rat; nucleic acid sequence; protein structure function; steroid hormone; sulfated glycoprotein 2; testis; tissue /cell culture; transcription factor; western blottings

The long-term goal of this research project is to delineate the molecular mechanisms involved in controlling male reproduction. One of the major advances we have made recently is to provide the first evidence that GATA-1 factor, a GATA-binding protein which was originally considered as an erythroid cell-specific transcription factor, plays an important role in regulating the transcription of testicular genes including inhibin/activin alpha- and beta-B-subunit genes in MA-10, a mouse Leydig tumor cell line. We also demonstrated that another testicular GATA-binding protein, GAT-4, can selectively transactivate inhibin/activin beta-B-subunit gene. In view that the mechanisms by which GATA-binding protein(s) interacts and transactivates testicular genes have never been investigated, in this application we propose to use inhibin/activin alpha- and beta-B- subunit genes as mosel systems to study the actions of GATA-1 and GATA-4 on the control of the expression of specific genes in the testis. Specific Aim 1 is to investigate the mechanisms by which GATA-1 interacts and transactivates the testicular rat inhibin alpha- subunit gene in MA-10 and rat Sertoli cells, to examine the functional domains of GATA-1, and to identify protein factors that confer the testis-specific transactivaion of the alpha-subunit gene by GATA-1. Specific Aim 2 is to compare the mechanisms by which GATA-1 and GATA-4 upregulate the promoter for. the 4.8-kb rat inhibin/activin beta-B-subunit gene transcript, to identify the GATA and GATA-like sequences in the beta-B-subunit promoter that are responsible for the transactivation by GATA-1 and/or GATA-4 factor in testicular cells, and to determine the functional domains of GATA-4 that interact and transactivate the promoter for the 4.8-kb beta-B-subunit transcript and compare with those of GATA-1 factor. Specific Aim 3 is to investigate the developmental and hormonal regulation of GATA- binding proteins in rat testis and compare to those previously observed in inhibin/activin alpha- and beta-B-subunit genes. Specific Aim 4 is to isolate and characterize new testicular GATA-binding protein(s) that transactivates alpha- and/or beta-B-subunit promoters. The observations obtained from the proposed studies may yield some new information on the delineation of the mechanisms involved in the controls of testicular physiology.

该研究项目的长期目标是描绘 参与控制男性生殖的分子机制。 一 我们最近取得的主要进展之一是提供了第一个 有证据表明 GATA-1 因子是一种 GATA 结合蛋白 最初被认为是红细胞特异性转录因子， 在调节睾丸转录中发挥重要作用 基因，包括抑制素/激活素α-和β-B-亚基基因 MA-10，小鼠 Leydig 肿瘤细胞系。 我们还证明了 另一种睾丸 GATA 结合蛋白 GAT-4 可以选择性地 反式激活抑制素/激活素β-B-亚基基因。 鉴于 GATA 结合蛋白相互作用的机制 反式激活睾丸基因从未被研究过，在这方面 应用我们建议使用抑制素/激活素α-和β-B- 亚基基因作为 mosel 系统来研究 GATA-1 的作用和 GATA-4对特定基因表达的控制 睾丸。 具体目标 1 是研究其机制 GATA-1 与睾丸大鼠抑制素 α-相互作用并反式激活 MA-10 和大鼠支持细胞中的亚基基因，以检查功能 GATA-1 结构域，并鉴定赋予 GATA-1 结构域的蛋白质因子 GATA-1 对 α 亚基基因的睾丸特异性反式激活。 具体目标 2 是比较 GATA-1 和 GATA-4 上调启动子。 4.8 kb 大鼠抑制素/激活素 β-B-亚基基因转录本，用于鉴定 GATA 和 GATA 样基因 β-B-亚基启动子中的序列负责 睾丸细胞中 GATA-1 和/或 GATA-4 因子的反式激活， 并确定 GATA-4 相互作用的功能域 反式激活 4.8 kb β-B 亚基转录物的启动子 并与GATA-1因子进行比较。 具体目标 3 是 研究 GATA-的发育和激素调节 大鼠睾丸中的结合蛋白并与之前的比较 在抑制素/激活素α-和β-B-亚基基因中观察到。具体的 目标 4 是分离和表征新的睾丸 GATA 结合 反式激活 α 和/或 β-B 亚基启动子的蛋白质。 从拟议研究中获得的观察结果可能会产生一些结果 关于界定所涉机制的新信息 睾丸生理学的控制。