MOLECULAR GENETICS OF SEX DETERMINATION IN C ELEGANS

如何调节细胞分化为一种细胞类型，而不是 再来一杯？如何协调调节细胞群以采用 同样的命运？我们建议解决这些基本问题： 后生动物发展通过分析控制命运的一个重大决定 线虫中的线虫：性命运的决定。 在过去的几年里，我们已经为实验奠定了基础 建议。首先，我们分析了tra-2基因及其rna产物。 在分子水平上。这项工作表明tra-2起着关键作用。 通过调节细胞通讯在蠕虫性别决定中的作用 通过调控细胞命运的转录调控因子。第二，我们有 对FOG-1基因进行了遗传分析，发现FOG-1是一个末端 生殖系中的性命调控者。 在未来五年，我们建议研究这些机制。 通过tra-2促进整个动物的雌性发育和 FOG-1通过它来指定生殖系中的精子发生。首先，我们 将描述单独的tra-2产物在性行为中的作用 决心。为此，我们将分析表型 表达三种tra-2 RNA中每一种的转基因动物。 第二，我们将提高针对tra-2蛋白的抗体，阐明它们的 细胞位置，并在整个发育过程中描述它们的特征 每种性别。第三，我们将测试预测的膜 蛋白pTra2A可能是一种受体。为此，我们将分析 Tra-2的自主性，或者使用组织特异性启动子驱动 TRA-2在转基因动物特定细胞中的表达 经典的马赛克分析。第四，我们将探索母体的作用 TrA-2在早期胚胎性别决定中的应用 遗传母体效应试验和RNA注射试验。第五, 我们将确定tra-2蛋白中介导HER-1的结构域 控制和促进女性发展，通过调节一个或多个 有限元基因。第六，我们将研究翻译规则的翻译。 2RNA的3‘端非编码区。第七，我们将克隆FOG-1基因、序列 FOG-1cDNA，鉴定和鉴定FOG-1转录本。这些 研究将阐明tra-2和fog-1的作用机制 这两个基因通过什么来调节和发挥它们的关键作用 决定性生活的命运。 这项工作与健康的相关性源于它对 理解细胞命运的基本控制，包括细胞 交流以及转录和翻译的调节。这些 控制是所有后生动物共有的，包括人类。

How are cells regulated to differentiate as one cell type rather than another? How are groups of cells coordinatedly regulated to adopt the same fate? We propose to approach these fundamental problems of metazoan development by analyzing the control of a major fate decision in the nematode Caenorhabditis elegans: determination of sexual fate. Over the past few years, we have laid the groundwork for experiments proposed. First, we have analyzed the tra-2 gene and its RNA products at the molecular level. This work suggests that tra-2 plays a pivotal role in worm sex determination by mediating cell communication and also by regulating transcriptional regulators of cell fate. Second, we have analyzed the fog-1 gene genetically and found that fog-1 is a terminal regulator of sexual fate in the germ line. During the next five years, we propose to investigate the mechanisms by which tra-2 promotes female development throughout the animal and by which fog-1 specifies spermatogenesis in the germ line. First, we will delineate the roles of the individual tra-2 products in sex determination. To this end, we will analyze the phenotypes of transgenic animals that express one of each of the three tra-2 RNAs. Second, we will raise antibodies to the tra-2 proteins, elucidate their cellular locations, and characterize them throughout development in each sex. Third, we will test the idea that the predicted membrane protein pTra2A may serve as a receptor. To this end, we will assay the autonomy of tra-2, either using tissue-specific promoters that drive tra-2 expression in particular cells of transgenic animals or by classic mosaic analysis. Fourth, we will explore the role of maternal tra-2 in early embryonic sex determination, using a combination of genetic maternal effect tests and RNA injection experiments. Fifth, we will identify domains in the tra-2 protein that mediate her-1 control and that promote female development by regulating one or more fem genes. Sixth, we will investigate translational regulation of tra- 2 RNA by its 3'UTR. Seventh, we will clone the fog-1 gene, sequence fog-1 cDNAs, and identify and characterize the fog-1 transcript. These studies will shed light on the mechanisms by which tra-2 and fog-1 are regulated and by which these two genes fulfill their key roles in determination of sexual fate. The health relatedness of this work derives from its contribution to an understanding of fundamental controls of cell fate, including cell communication and transcriptional and translational regulation. These controls are common to all metazoans, including man.