MOLECULAR BASIS FOR AUTONOMOUS CELL FATES IN C ELEGANS

线虫自主细胞命运的分子基础

• 批准号: 2186003
• 负责人: JAMES R PRIESS
• 金额: $ 25.15万
• 依托单位: FRED HUTCHINSON CANCER RESEARCH CENTER
• 依托单位国家: 美国
• 财政年份: 1993
• 资助国家: 美国
• 起止时间: 1993-01-01 至 1996-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2186003
• 关键词: Caenorhabditis elegans; DNA binding protein; alleles; cell differentiation; developmental genetics; early embryonic stage; embryo /fetus cell /tissue; embryogenesis; fluorescence microscopy; gel mobility shift assay; gene expression; gene interaction; gene mutation; genetic mapping; in situ hybridization; light microscopy; nucleic acid probes; nucleic acid sequence; phenotype; polymerase chain reaction; protein structure function; video recording system

We want to understand how embryonic cells become committed to particular pathways of differentiation. Molecular and genetic analysis of development in the fruit fly Drosophila and the nematode Caenorhabditis elegans already has resulted in the identification of several key genes involved in determining cell fate. The vertebrate homologs of many of these genes have now been identified, and mutations in some of these homologous genes are associated with tumor formation. Analysis of the function of these genes in model systems like flies and nematodes is likely to provide a better understanding of how the homologous genes function in vertebrates, and possibly suggest therapeutic approaches to correcting functional defects. Although substantial progress has been made in understanding the very early events in the Drosophila embryo, almost nothing is known about the molecules that control early embryogenesis in C. elegans. Because the early development of these two organisms is very different, analysis of embryogenesis in C. elegans should provide a broader understanding of animal development. We have used a genetic approach to identify a small set of genes in C. elegans that appear to play critical roles in determining the fate of early embryonic cells. These genes are named skn-1; skn-2; pie-1; mex-1 and mex-3. The general aim of the proposed work is the molecular analysis of each of these genes and the localization of their gene products in the embryo. skn-1 encodes a protein with a sequence similarity to the DNA-binding domain of a set of transcription factors called BZIP proteins. However in overall structure the skn-1 protein does not resemble BZIP family members and appears to be a novel protein. Experiments are proposed to determine if skn-1 can bind specific DNA sequences, and to identify genes whose products interact with skn-1 to control cell fate. Genetic analysis of pie-1 and skn-2 mutants suggests the products of these genes may interact. Genetic experiments are proposed to identify alleles of pie-1 that will be useful for molecular analysis, as well as other genes that may interact with pie-1. pie-1 and skn-2 mutants lack a particular cell type that mex-3 mutants overproduce. Analysis of mex-3 mutants should provide a better insight into how pie-1 and skn-2 function, and experiments are proposed to complete a phenotypic characterization of the mex-3 embryonic defect.

我们想了解胚胎细胞是如何 分化的途径。 分子和遗传分析 果蝇和小杆线虫的发育 已经鉴定出了几个关键基因 参与决定细胞的命运。 许多脊椎动物的同源物 这些基因现在已经被鉴定出来，其中一些基因的突变 同源基因与肿瘤形成有关。 分析 这些基因在苍蝇和线虫等模型系统中的功能是 这可能会让我们更好地理解同源基因 功能，并可能提出治疗方法， 纠正功能缺陷。 尽管在理解这一问题上取得了实质性进展， 果蝇胚胎的早期事件，几乎没有什么是已知的， 控制C.优雅的 因为 这两种生物的早期发育是非常不同的，分析 C.胚胎发生优雅应该提供更广泛的理解， 动物发展。 我们用遗传学的方法鉴定了一个小的 C.这些线虫似乎在 决定早期胚胎细胞的命运。 这些基因被命名为 skn-1; skn-2; pie-1; mex-1和mex-3。 建议的总体目标 工作是对这些基因中的每一个进行分子分析， 基因产物在胚胎中的定位。 skn-1编码一种 与一组DNA结合结构域具有序列相似性的蛋白质， 称为BZIP蛋白的转录因子。 但在整体结构上 skn-1蛋白与BZIP家族成员不同， 是一种新的蛋白质。 提出了实验来确定skn-1是否可以 结合特定的DNA序列，并确定其产物 与skn-1相互作用以控制细胞命运。 pie-1基因的遗传分析， skn-2突变体表明这些基因的产物可能相互作用。 遗传 提出了实验来鉴定pie-1的等位基因， 用于分子分析，以及其他可能与 派-1。 pie-1和skn-2突变体缺乏mex-3 突变体会过度繁殖 对mex-3突变体的分析应该提供一个更好的 深入了解pie-1和skn-2的功能，并提出实验 以完成MEX-3胚胎缺陷的表型表征。