Messenger RNA localization in Drosophila oogenesis

果蝇卵子发生中的信使 RNA 定位

• 批准号: 6457559
• 负责人: EDWIN C STEPHENSON
• 金额: $ 13.75万
• 依托单位: UNIVERSITY OF ALABAMA IN TUSCALOOSA
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-03-01 至 2005-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6457559
• 关键词: Drosophilidae; cytoskeleton; dynein ATPase; egg /ovum; fluorescence microscopy; gene mutation; green fluorescent proteins; in situ hybridization; messenger RNA; mutant; nucleic acid quantitation /detection; oogenesis; protein localization; protein protein interaction; protein structure function; yeasts

DESCRIPTION: (provided by applicant) The subcellular localization of messenger RNAs is a widespread mechanism for localizing cytoplasmic proteins in cells. This proposal addresses the mechanism by which localized mRNAs are anchored in a specific subcellular region, a process that probably involves the attachment of RNA-protein complexes to the cytoskeleton. In the model system under consideration in this proposal, two mRNAs, bicoid and htsN4 are each localized at the anterior margin of Drosophila oocytes. It has been proposed that the swallow protein acts to link the mRNAs to the microtubule cytoskeleton, perhaps through the microtubule motor molecule dynein. The experiments proposed here will investigate several features of this model: Hypothesis 1: The dynein motor complex plays an essential role in anchoring bicoid and htsN4 mRNAs at the anterior oocyte margin. This idea will be tested by determining whether dynein mutants affect bicoid/htsN4 mRNA localization. Hypothesis 2: The swallow protein is associated with the dynein motor complex and mediates RNA localization through this association. This idea will be tested by determining the subcellular localization and co-localization of the swallow protein, determining the effects of dynein mutants on swa protein localization, and by identifying and mapping interacting domains in proteins that associate with swallow. Hypothesis 3: The effects of swallow mutations on cytoskeletal organization are indirect, caused by the failure to localize htsN4 mRNA and the adducin-like protein it encodes. Swallow mutant embryos have defects in the cytoskeleton that have confused past attempts to understand the role of swallow in development. Here we test the idea that htsN4 mRNA and protein mislocalization are sufficient to account for the observed cytoskeletal defects, and that swa only has an indirect role in cytoskeletal organization.

描述：（由申请人提供）Messenger的亚细胞定位 RNA是将细胞质蛋白定位在细胞中的广泛机制。 该提案解决了局部mRNA锚定在内的机制 特定的亚细胞区域，可能涉及附件的过程 RNA蛋白复合物的细胞骨架。在模型系统中 在此提案中的考虑，两个mRNA，BICOID和HTSN4都位于本地化 在果蝇卵母细胞的前缘。有人提出 吞咽蛋白的作用是将mRNA与微管细胞骨架联系起来，也许 通过微管运动分子动力蛋白。这里提出的实验 将研究该模型的几个功能： 假设1：动力蛋白运动复合物在锚定方面起着至关重要的作用 前卵母细胞边缘的双聚体和HTSN4 mRNA。这个想法将被测试 通过确定动力蛋白突变体是否影响双叶素/HTSN4 mRNA定位。 假设2：吞咽蛋白与动力蛋白运动复合物有关 并通过这种关联介导RNA定位。这个想法将是 通过确定细胞亚定位和共定位的测试 吞咽蛋白，确定动力蛋白突变体对SWA蛋白的影响 通过定位，并通过识别和映射蛋白质中的相互作用域 那个与燕子相关。 假设3：燕子突变对细胞骨架组织的影响是 间接，是由于未能定位HTSN4 mRNA和附加素样引起的 蛋白质它编码。燕子突变胚胎在细胞骨架中存在缺陷 过去的尝试使吞咽作用的尝试困惑 发展。在这里，我们测试了HTSN4 mRNA和蛋白质错误定位的想法 足以说明观察到的细胞骨架缺陷，而SWA 仅在细胞骨架组织中具有间接作用。