GERM CELL DIFFERNTIATION IN DROSOPHILA

果蝇生殖细胞分化

• 批准号: 6194373
• 负责人: DENNIS M MCKEARIN
• 金额: $ 29.9万
• 依托单位: UT SOUTHWESTERN MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 1991
• 资助国家: 美国
• 起止时间: 1991-05-01 至 2004-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6194373
• 关键词: Drosophilidae; alleles; cell cell interaction; cell differentiation; cell growth regulation; cell type; cytogenetics; developmental genetics; fertility; gene induction /repression; gene interaction; gene mutation; genetic transcription; immunoprecipitation; molecular cloning; molecular genetics; nucleic acid sequence; oogenesis; protein structure function; stem cells; yeast two hybrid system

DESCRIPTION (Adapted from investigator's abstract): The goal of the proposed experiments is to discover the mechanism by which the bag-of-marbles (bam) and benign gonial cell neoplasm (bgcn) gene products act to control the earliest steps in female germ cell differentiation from germ line stem cell precursors. Mutations in bam and bgcn have the same phenotype. Loss of function of either gene causes over proliferation of what appear to be primordial germ cells or germ line stem cells in females and over proliferation of interconnected, mitotically amplifying spermatogonia in males. Thus, in both sexes, Bam and Bgcn play a critical role in choices between proliferations as an undifferentiated precursor versus initiation of differentiation. The investigator proposes four aims to explore how Bam and Bgcn function. 1) To determine if the bam and bgcn proteins act together, the investigator will determine if bam and bgcn proteins co-localize in the cell or co-immunoprecipitate. 2) To investigate the mechanism of action of Bgcn and Bam, especially a possible role Bgcn in translational control, the investigator will screen for proteins that interact with Bgcn in the yeast two hybrid system and test if the Bgcn protein binds to RNA (as predicted based on its sequence homology). 3) To explore whether Bam protein function in the fusome is important for regulation of early germ cell differentiation, the investigator will identify the fusome targeting sequences in the Bam protein. If these can be eliminated without destroying Bam activity, it would argue against the investigator's model that localization of Bam protein to the fusome is important for its function in controlling germ cell fate. 4) To identify other components of the mechanism, the investigator will screen for mutations that enhance a weak bam mutant. In addition, the investigator will attempt to identify upstream regulations of bam using a screen for mutations that alter the cell type specific pattern of bam transcription or tiotranslantion.

描述（改编自研究者摘要）：拟定的目标 实验是为了发现机制，其中袋的大理石（BAM）和 良性生殖腺细胞肿瘤（bgcn）基因产物的作用是控制最早的 从生殖系干细胞前体向雌性生殖细胞分化的步骤。 bam和bgcn的突变具有相同的表型。功能丧失 基因导致原始生殖细胞过度增殖， 生殖系干细胞在女性和过度增殖的相互联系， 雄性的精原细胞有丝分裂扩增。因此，在两性中，巴姆和 Bgcn在作为一个 未分化的前体与分化的起始。的 研究者提出了四个目标来探索Bam和Bgcn如何发挥作用。 1)为了确定bam和bgcn蛋白是否一起起作用， 将确定BAM和BGCN蛋白是否共定位在细胞中， 免疫共沉淀。 2)探讨Bgcn和Bam的作用机制，特别是 Bgcn在翻译控制中的可能作用，研究者将筛选 在酵母双杂交系统中与Bgcn相互作用的蛋白质， Bgcn蛋白与RNA结合（根据其序列同源性预测）。 3)为了探讨Bam蛋白在融合体中的功能是否对 早期生殖细胞分化的调节，研究者将确定 Bam蛋白中的融合体靶向序列。如果这些都能被消除 在不破坏Bam活动的情况下，它会反驳调查人员的 Bam蛋白在融合体中的定位对于其在融合体中的表达是重要的。 控制生殖细胞命运的功能。 4)为了确定该机制的其他组成部分，研究人员将筛选 增强弱BAM突变体的突变。此外，调查员 将尝试使用屏幕识别bam的上游规则， 改变BAM转录的细胞类型特异性模式的突变， 翻译