GATA-BINDING PROTEINS REGULATE TESTICULAR GENE EXPRESSION

GATA 结合蛋白调节睾丸基因表达

• 批准号: 6108355
• 负责人: CHING-LING C CHEN
• 金额: $ 23.88万
• 依托单位: POPULATION COUNCIL
• 依托单位国家: 美国
• 财政年份: 1999
• 资助国家: 美国
• 起止时间: 1999-04-01 至 2000-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6108355
• 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述 与控制男性繁殖有关的分子机制。 一 我们最近取得的主要进步是提供第一个 证据表明GATA-1因子是一种GATA结合蛋白，是 最初被认为是红细胞特异性转录因子， 在调节睾丸的转录中起着重要的作用 包括抑制蛋白/活化素α-和β-b亚基基因在内的基因 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可以有选择地 反式激活抑制素/活化素β-B-亚基基因。 考虑到 GATA结合蛋白相互作用的机制和 从未研究过睾丸基因的反式激活。 我们建议使用抑制素/激活素α-和beta-b- 亚基基因作为Mosel系统研究GATA-1和 GATA-4关于控制特定基因表达的GATA-4 睾丸。 具体目的1是调查该机制 GATA-1相互作用并反式激活睾丸大鼠抑制剂α- MA-10和大鼠Sertoli细胞中的亚基基因，以检查功能 GATA-1的域，并确定赋予蛋白质因子 GATA-1对α-亚基基因的睾丸特异性反抗性。 具体目标2是比较GATA-1和GATA-1和 GATA-4上调了启动子。 4.8-kb大鼠抑制素/激活素 beta-b-subunit基因转录本，以识别gata和gata样 beta-b-subunit启动子中负责的序列 睾丸细胞中GATA-1和/或GATA-4因子的反式激活， 并确定相互作用的GATA-4的功能域 对4.8-kb beta-b-Subunit转录本的启动子进行反式激活 并与GATA-1因子相比。 具体目标3是 研究GATA-的发育和激素调节 在大鼠睾丸中结合蛋白质，并与先前的蛋白质进行比较 在抑制蛋白/活化素α-和β-b-subunit基因中观察到。具体的 AIM 4是隔离和表征新的睾丸GATA结合 反式激活α-和/或β-b-subunit启动子的蛋白质。 从提出的研究中获得的观察结果可能会产生一些 有关描述所涉及的机制的新信息 睾丸生理的控制。