PROTO-ONCOGENES IN GAMETOGENESIS AND EARLY DEVELOPMENT
PROTO-ONCOGENES IN GAMETOGENESIS AND EARLY DEVELOPMENT
批准号:
2378511
负责人:
GEOFFREY M COOPER
金额:
$23.53万
依托单位:
依托单位国家:
美国
项目类别:
财政年份:
1996
资助国家:
美国
项目状态:
已结题
起止时间:
1996-03-01 至 1998-02-28
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
中文摘要
该项目的总体目标是了解原型的作用,
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,
在减数分裂中起关键作用的c-mos基因的转录
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,导致其在雄性中特异性表达
和女性生殖细胞。对c-mos的这种监管不仅需要
在生殖细胞中实现适当的c-mos表达,
体细胞中的不适当表达,这可能导致
细胞死亡或肿瘤转化。 值得注意的是,C-
mos表达在胚胎发生早期下调,这可能是
防止胚胎分裂停滞,
Mos作为细胞生长抑制因子的作用。
c-mos在体细胞中的转录被一个负性的
调控元件(NRE)上游的c-mos启动子。 此外,本发明还提供了一种方法,
最近的研究已经确定了一种候选体细胞阻遏物,
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
抑制c-mos转录的作用机制,
其他生殖细胞特异性基因。
c-mos在卵母细胞中的表达不受NRE的影响,
仅需要最少的启动子,包括起始子(Inr)元件。
这些发现提示了c-mos在卵母细胞中转录的假说
这是由高水平的基础转录活性引起的,
抑制在两细胞胚胎中。 这将进一步调查
c-mos Inr基因转录活性的研究
卵母细胞和胚胎,连同表达分析,
c-mos阻遏物在胚胎发育过程中的功能。
拟议的研究有三个具体目标:
1. 一个体细胞cDNA克隆的分离和鉴定
c-mos转录的阻遏物。
2. c-mos的结构/功能分析及机理研究
阻遏作用
3. 卵母细胞c-mos转录调控的研究
早期胚胎
英文摘要
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.
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