PROTO-ONCOGENES IN GAMETOGENESIS AND EARLY DEVELOPMENT

配子发生和早期发育中的原癌基因

• 批准号: 2883127
• 负责人: GEOFFREY M COOPER
• 金额: $ 24.88万
• 依托单位: BOSTON UNIVERSITY (CHARLES RIVER CAMPUS)
• 依托单位国家: 美国
• 财政年份: 1998
• 资助国家: 美国
• 起止时间: 1998-03-01 至 2002-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2883127
• 关键词: 3T3 cells; DNA binding protein; DNA footprinting; HeLa cells; affinity chromatography; developmental genetics; early embryonic stage; embryo /fetus; gametogenesis; gel mobility shift assay; gene expression; genetic promoter element; genetic regulatory element; laboratory mouse; mammalian embryology; northern blottings; nucleic acid probes; oogenesis; protein purification; protein structure function; protooncogene; spermatogenesis; transcription factor; western blottings

The overall goal of the project is understanding the roles of proto- oncogenes in gametogenesis and early development of mammalian embryos. The proposed research is focused on the mechanisms that regulate transcription of the c-mos gene, which plays a critical role in meiosis of vertebrate oocytes. Transcription of c-mos is subject to stringent tissue--specific regulation, leading to its specific expression in male and female germ cells. Such regulation of c-mos is needed not only to achieve appropriate c-mos expression in germ cells but also to prevent inappropriate expression in somatic cells, which can result in either cell death or neoplastic transformation. It is also noteworthy that c- mos expression is downregulated early in embryogenesis, which may be necessary to prevent arrest of embryonic cleavage divisions resulting from the action of Mos as a cytostatic factor. Transcription of c-mos in somatic cells is suppressed by a negative regulatory element (NRE) upstream of the c-mos promoter. In addition, recent studies have identified a candidate somatic cell repressor that binds to a functional element within the c-mos NRE. We now plan to isolate a cDNA clone of the repressor protein and to characterize its mechanism of action in suppressing transcription of c-mos, and possibly other germ cell-specific genes, in somatic cells. Expression of c-mos in oocytes, which is not affected by the NRE, requires only a minimal promoter, including an initiator (Inr) element. These findings suggest the hypothesis that c-mos transcription in oocytes results from a high level of basal transcription activity, which is then suppressed in two-cell embryos. This will be investigated by further studies of the activity of the c-mos Inr in directing transcription in oocytes and embryos, together with analysis of the expression and function of the c-mos repressor during embryonic development. The proposed studies have three specific aims: 1. Isolation and characterization of a cDNA clone for a somatic cell repressor of c-mos transcription. 2. Structure/function analysis and studies of the mechanism of c-mos repressor action. 3. Investigation of the regulation of c-mos transcription in oocytes and early embryos.

该项目的总体目标是了解原型的角色 配子发生和哺乳动物胚胎早期发育中的癌基因。 拟议的研究重点是调节机制 c-mos 基因的转录，在减数分裂中发挥关键作用 脊椎动物的卵母细胞。 c-mos的转录受到严格的 组织特异性调节，导致其在男性中特异性表达 和女性生殖细胞。对 c-mos 的这种调节不仅需要 在生殖细胞中实现适当的 c-mos 表达，同时还可以防止 体细胞中的不适当表达，可能导致 细胞死亡或肿瘤转化。 还值得注意的是，c—— mos 表达在胚胎发生早期下调，这可能是 防止胚胎卵裂分裂所必需的 来自Mos作为细胞抑制因子的作用。 体细胞中 c-mos 的转录受到负性抑制 c-mos 启动子上游的调控元件（NRE）。 此外， 最近的研究发现了一种候选体细胞抑制因子 与 c-mos NRE 内的功能元件结合。 我们现在计划 分离阻遏蛋白的 cDNA 克隆并表征其特征 抑制c-mos转录的作用机制，以及可能 体细胞中的其他生殖细胞特异性基因。 卵母细胞中c-mos的表达，不受NRE影响， 仅需要一个最小的启动子，包括一个引发子（Inr）元件。 这些发现提出了这样的假设：卵母细胞中的 c-mos 转录 来自高水平的基础转录活性的结果，然后 在双细胞胚胎中受到抑制。 此事将进一步调查 c-mos Inr 在指导转录中的活性研究 卵母细胞和胚胎，以及表达和分析 c-mos 阻遏蛋白在胚胎发育过程中的功能。 拟议的研究有三个具体目标： 1. 体细胞 cDNA 克隆的分离和表征 c-mos转录抑制子。 2. c-mos结构/功能分析及机理研究 抑制作用。 3. 卵母细胞c-mos转录调控的研究 和早期胚胎。

项目成果

c-mos expression in mouse oocytes is controlled by initiator-related sequences immediately downstream of the transcription initiation site.

小鼠卵母细胞中的 c-mos 表达由紧邻转录起始位点下游的起始子相关序列控制。

• DOI: 10.1128/mcb.11.10.5190-5196.1991
• 发表时间: 1991
• 期刊: Molecular and cellular biology
• 影响因子: 5.3
• 作者: Pal,SK;Zinkel,SS;Kiessling,AA;Cooper,GM
• 通讯作者: Cooper,GM

Identification of a negative regulatory element that inhibits c-mos transcription in somatic cells.

鉴定抑制体细胞中 c-mos 转录的负调控元件。

• DOI: 10.1128/mcb.12.5.2029-2036.1992
• 发表时间: 1992
• 期刊: Molecular and cellular biology
• 影响因子: 5.3
• 作者: Zinkel,SS;Pal,SK;Szeberényi,J;Cooper,GM
• 通讯作者: Cooper,GM

Functional analysis of sperm from c-mos(-/-) mice.

c-mos(-/-) 小鼠精子的功能分析。

• DOI: 10.1002/mrd.10140
• 发表时间: 2002
• 期刊: Molecular reproduction and development.
• 作者: Gross,VeraS;Cooper,GeoffreyM
• 通讯作者: Cooper,GeoffreyM

Identification of a candidate c-mos repressor that restricts transcription of germ cell-specific genes.

鉴定限制生殖细胞特异性基因转录的候选 c-mos 阻遏蛋白。

• DOI: 10.1128/mcb.15.10.5369
• 发表时间: 1995
• 期刊: Molecular and cellular biology
• 影响因子: 5.3
• 作者: Xu,W;Cooper,GM
• 通讯作者: Cooper,GM

The c-mos gene product is required for cyclin B accumulation during meiosis of mouse eggs.

c-mos 基因产物是小鼠卵减数分裂过程中细胞周期蛋白 B 积累所必需的。

• DOI: 10.1073/pnas.88.17.7869
• 发表时间: 1991
• 期刊: Proceedings of the National Academy of Sciences of the United States of America
• 影响因子: 11.1
• 作者: O'Keefe,SJ;Kiessling,AA;Cooper,GM
• 通讯作者: Cooper,GM