Molecular Mechanisms of Adhesion in Dictyostelium

盘基网柄菌粘附的分子机制

• 批准号: 9218909
• 负责人: William Loomis
• 金额: $ 33万
• 依托单位: University of California-San Diego
• 依托单位国家: 美国
• 项目类别: Continuing Grant
• 财政年份: 1993
• 资助国家: 美国
• 起止时间: 1993-02-01 至 1997-01-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=9218909&HistoricalAwards=false
• 关键词: Molecular; Mechanisms; Adhesion; Dictyostelium

Three independent mechanisms of cell-cell adhesion function during development of Dictyostelium discoideum. The early adhesion system is mediated by a small protein, gp24, that first appears on the surface of cells several hours after initiation of development and preferentially accumulates in prestalk cells. There are three closely linked genes, csbA, csbB, and csbC, encoding gp24 that appear to be functionally redundant. Transformed cell lines in which one or two of these genes are deleted show alterations in early adhesion. Moreover, cells lacking csbC culminate only after a considerable delay and form gnarled fruiting bodies. Dr. Loomis will use homologous recombination to replace all three csb loci with a transformation vector carrying the pyr5-6 gene. The phenotype of the resulting strains will be analyzed in detail to determine the consequences to cell-cell adhesion and subsequent development. He will genetically rescue these triple deletion strains by transformation with a wild-type copy of one or more of the csb genes and then explore the molecular mechanism of adhesion by determining the functionality of variants generated by site specific mutagenesis. Using Restriction Enzyme Mediated Integration (REMI) to tag developmentally important genes, he will isolate and characterize other genes essential for cell-cell adhesion during early development.

细胞间粘附功能的三种独立机制 盘基网柄藻的发育。早期粘附系统 是由一种小蛋白gp 24介导的，它首先出现在 在发育开始后几小时， 优先在前柄细胞中积累。有三 紧密相连的基因csbA、csbB和csbC，编码gp 24， 似乎是多余的转化细胞系 这些基因中的哪一个或哪两个被删除， 早期粘连此外，缺乏csbC的细胞只有在 一个相当大的延迟和形成多节的子实体。卢米斯博士 将使用同源重组替换所有三个csb基因座 用携带pyr 5 -6基因的转化载体转化。的 将详细分析所得菌株的表型， 确定对细胞间粘附和随后的 发展他将从基因上拯救这些三重缺失 通过用以下一种或多种的野生型拷贝转化菌株 csb基因，探讨黏附的分子机制 通过确定站点生成的变体的功能， 特异性诱变使用限制酶介导的整合 （REMI）标记发育重要的基因，他将分离， 表征细胞间粘附所必需的其他基因， 早期发展。