TF IIIA GENE EXPRESSION IN OOCYTES AND SOMATIC CELLS

卵母细胞和体细胞中的 TF IIIA 基因表达

• 批准号: 2179703
• 负责人: WILLIAM L TAYLOR
• 金额: $ 14.14万
• 依托单位: UNIVERSITY OF TENNESSEE HEALTH SCI CTR
• 依托单位国家: 美国
• 财政年份: 1988
• 资助国家: 美国
• 起止时间: 1988-07-01 至 1996-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2179703
• 关键词: DNA binding protein; DNA footprinting; RNA splicing; Xenopus oocyte; cell transformation; developmental genetics; gene expression; genetic promoter element; genetic regulatory element; laboratory rabbit; messenger RNA; molecular cloning; mutant; nucleic acid probes; nucleic acid sequence; oogenesis; polymerase chain reaction; posttranslational modifications; transcription factor

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.

描述（改编自申请人的摘要）： 这项工作是为了了解分子机制， 转录因子IIIA（TFIIIA）基因是发育调控的。 的 迄今为止的研究主要集中在顺式元件的鉴定上 TFIIIA基因在未成熟卵母细胞、成熟卵母细胞和成熟卵母细胞中的表达所需 卵母细胞和体细胞。 这些顺式元件中的八个或更多个已经被 其中一些似乎是发育调节的。 的 发育调控元件在不同的细胞中功能性表达， 有时，一种是特异性的未成熟卵母细胞和其他（几个 阴性和一个阳性）是“胚胎特异性的”（在胚胎中不起作用）。 卵母细胞）。 这些因素被调节的时间对应于 当TFIIIA mRNA水平变化时（TFIIIA 10倍以上）， mRNA在未成熟卵母细胞比成熟卵母细胞和大约10-5 TFIIIA mRNA在成熟卵母细胞中的表达高于在原肠胚后胚胎中的表达）。的 研究人员已经鉴定出与大多数细胞结合的反式作用因子， 这些元件，并获得了推定的cDNA克隆之一， 发育调节因子（与不成熟的 卵母细胞特异性元件）。 本提案的重点是扩大关于相互作用的结果 这些因素之间的进一步表征的因素， 绑定到这些元素。 研究人员建议将各种 反式作用因子和/或分离编码这些因子的cDNA克隆。 这些cDNA克隆将提供一个分子探针， 因素 这些因素的作用及其相互作用， 然后将使用体内和体外方法检测其它转录因子。 体外方法。 质量和数量上的差异 将确定这些因素的结构、活性和相互作用 在发育过程中的不同时期