TFIIIA GENE EXPRESSION IN OOCYTES & SOMATIC CELLS

描述(改编自申请人的摘要)：的主要目标 这项工作是为了了解分子机制，通过 转录因子IIIA(TFIIIA)基因受发育调控。这个 迄今为止的研究主要集中在顺式元素的鉴定上。 TFIIIA基因在未成熟卵母细胞、成熟卵母细胞中表达所需 卵母细胞和体细胞。这些顺式元素中有八个或更多已经 其中几个似乎受发育调节。这个 发育调节元件在不同的功能上表达 一个是针对未成熟卵母细胞的，另一个是(几个) 阴性和一种阳性)是“胚胎特有的”(在 卵母细胞)。这些因素被调节的时间对应于 TFIIIA mRNA水平变化的发育时间(TFIIIA增加10倍 未成熟卵母细胞的mRNA量比成熟卵母细胞高出约10-5 TFIIIA mRNA在成熟卵母细胞中的表达高于原肠胚后)。这个 研究人员已经确定了与大多数 这些元件，并已获得其中一个假定的cdna克隆。 发育调节因子(与未成熟者结合的因子 卵母细胞特有的成分)。 这项提议集中于扩展关于相互作用的结果 在这些元素和进一步描述这些因素之间的关系 绑定到这些元素。调查员建议将各种不同的 反式作用因子和/或分离编码这些因子的cDNA克隆。 这些cdna克隆将为人们提供针对这些基因的分子探针。 各种因素。这些因素的作用以及它们与 其他转录因子将在体内和体内进行检测。 试管方法。在质量和数量上的差异 这些因素的结构、活性和相互作用将被确定 在发育过程中的不同时期。

DESCRIPTION (Adapted from the applicant's abstract): The primary goal of this work is to understand the molecular mechanism by which the Transcription Factor IIIA (TFIIIA) gene is developmentally regulated. The studies to date have concentrated on the identification of cis-elements required for the expression of the TFIIIA gene in immature oocytes, mature oocytes and somatic cells. Eight or more of these cis-elements have been identified, several of which appear to be developmentally regulated. The developmentally regulated elements are functionally expressed at different times, one is specific for immature oocytes and the others (several negative and one positive) are "embryo-specific" (not functional in oocytes). The times at which these factors are regulated correspond to times in development when TFIIIA mRNA levels change (10-fold more TFIIIA mRNA in immature oocytes than in mature oocytes and approximately 10-5 more TFIIIA mRNA in mature oocytes than in post-gastrula embryos). The investigator has identified trans-acting factors which bind to most of these elements and have obtained putative cDNA clones for one of the developmentally regulated factors (the one which binds to the immature oocyte-specific element). This proposal concentrates on expanding the results on the interactions between these elements and the further characterization of factors which bind to these elements. The investigator proposes to isolate the various trans-acting factors and/or isolate cDNA clones encoding these factors. These cDNA clones will then provide one with molecular probes for these factors. The functions of these factors and interactions between them and other transcription factors will then be examined using in vivo and in vitro approaches. Qualitative and quantitative differences in the structure, activity, and interactions of these factors will be determined at various times during development.