MAGO AND TSUNAGI FUNCTION IN DROSOPHILA OOGENESIS

MAGO 和 Tsunagi 在果蝇卵发生中的作用

• 批准号: 6386962
• 负责人: ROBERT E BOSWELL
• 金额: $ 29.22万
• 依托单位: UNIVERSITY OF COLORADO
• 依托单位国家: 美国
• 财政年份: 1999
• 资助国家: 美国
• 起止时间: 1999-05-01 至 2003-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6386962
• 关键词: Drosophilidae; biological signal transduction; cellular polarity; gene deletion mutation; gene expression; genetic mapping; green fluorescent proteins; immunoprecipitation; matrix assisted laser desorption ionization; molecular asymmetry; oogenesis; phenotype; point mutation; protein protein interaction; protein structure function; site directed mutagenesis

DESCRIPTION: (Adapted from investigator's abstract) The proposal focuses on the molecular mechanisms which establish asymmetries in the oocyte of Drosophila melanogaster. During oogenesis the posterior follicle cells send a signal of unknown nature to the developing oocyte. This signal leads to a polarization of the oocyte cytoskeleton, which then directs the localization of determinants such as oskar and bicoid RNA to the anterior and posterior poles of the egg. The investigator has previously identified a gene, mago nashi (mago) which plays a role in the signal transduction mechanism by which the oocyte receives the polarizing information from the posterior follicle cells. A second protein, Tsunami (Tsu) has been found to interact with Mago in yeast 2-hybrid screens. The proposal is aimed at providing insight into the in vivo function of both Mago and Tsu in Drosophila. The first goal of the proposal is to map domains of the Mago protein which are necessary for function and subcellular localization. Point mutations and deletions will be introduced into the Mago protein and the cellular distribution of these mutant proteins will be monitored. The protein will also be targeted to specific cellular locations, and the phenotypes of such artificially localized proteins will be determined. Tsunagi is a previously uncharacterized gene, and goal 2 of the proposal is the characterization of the role of tsu in development. Mutations in tsu will be searched for among the pre-existing mutants in the region. Such mutations will then be further characterized. The expression pattern of tsu RNA and protein will also be determined. Given that Tsu contains RNA binding motifs SELEX procedures will be performed in an attempt at defining RNA sequences that are bound by Tsu. If potential target proteins can be identified they will be further studied. The third goal of the proposal addresses the question whether Mago and Tsu interact with each other in vivo, and if these experiments are positive, the domains required for the interaction will be determined. (In the supplemental material that was submitted in September, the PI reports that Tsu and Mago indeed can indeed be co-immunoprecipitated from fly extracts). The fourth goal of the proposal involves screens for additional genes that act in the mago pathway. These screens have already been initiated and some promising candidates have been isolated, and will be further studied.

描述：(改编自调查人员摘要)该提案侧重于 果蝇卵母细胞不对称性形成的分子机制 黑猩猩。在卵子发生过程中，后卵泡细胞发出信号 对发育中的卵母细胞来说是未知的性质。该信号导致偏振 卵母细胞的细胞骨架，然后指导决定因素的定位 例如，Oskar和双体RNA到卵子的前后两极。 研究人员此前已经确定了一种基因，Mago nashi(MAGO)，它 在卵母细胞接收信号的信号传递机制中起作用 来自后毛囊细胞的极化信息。第二种蛋白质， 海啸(Tsu)已被发现在酵母双杂交筛选中与Mago相互作用。 该提案旨在提供对两者在体内功能的洞察力 《果蝇》中的Mago和Tsu。该提案的第一个目标是绘制 Mago蛋白是功能和亚细胞定位所必需的。 点突变和缺失将被引入到Mago蛋白和 这些突变蛋白的细胞分布将受到监测。这种蛋白质 也将针对特定的细胞位置，以及表型 这种人工定位的蛋白质将被确定。Tsunagi是一个 以前没有特征的基因，该提案的目标2是 描述技术工会在发展中的作用。TSU中的突变将是 在该地区已有的变种人中寻找。这样的突变将 然后进一步刻画。苏云金杆菌RNA和蛋白质的表达模式 也将被确定。鉴于Tsu包含RNA结合基序SELEX 将执行程序，以尝试定义符合以下条件的RNA序列 被Tsu捆绑。如果能鉴定出潜在的靶蛋白，它们就会 进一步研究。 提案的第三个目标解决了Mago和Tsu 在体内相互作用，如果这些实验是积极的， 将确定交互所需的域。(在补充资料中 9月份提交的材料，PI报告说，Tsu和Mago 确实可以从苍蝇提取物中进行免疫共沉淀)。第四个目标 该提案涉及对在MAGO中起作用的额外基因的筛选 路径。这些筛查已经启动，一些有希望的 候选人已被隔离，并将进行进一步研究。