Resolving the nuclear fusion steps during yeast mating

解决酵母交配过程中的核融合步骤

• 批准号: 7233370
• 负责人: PATRICIA G MELLOY
• 金额: $ 1.2万
• 依托单位: PRINCETON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-12-16 至 2006-08-25
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7233370
• 关键词: Saccharomyces cerevisiae; behavioral /social science research tag; cell cycle; electron microscopy; fluorescence microscopy; fungal proteins; green fluorescent proteins; membrane fusion; microorganism reproduction; nuclear membrane; polymerase chain reaction; postdoctoral investigator; protein structure function; protein transport; sex behavior

DESCRIPTION (provided by applicant): During the yeast mating process, nuclear fusion occurs between two haploid nuclei of the mating pairs, forming a diploid nucleus. Therefore, membrane fusion is a critical step in the completion of zygote formation, although little is known about this process. In the simplest model, the inner and outer membranes fuse simultaneously beginning at the membranous half-bridge of the spindle pole body. In the other model, nuclear fusion actually occurs in three steps: first the outer membranes fuse, then the inner membranes fuse, and finally spindle pole body fusion completes the process. To determine whether nuclear fusion occurs in one or three steps, we are generating CFP-and YFP-tagged nuclear, ER luminal, and integral membrane proteins to follow the temporal order of their transfer during nuclear fusion. We have begun our studies using a nuclear pore marker, Nic96p, as a nuclear fusion marker. Using Nic96-CFP/-YFP, we are able to establish the timing of the nuclear fusion event in live cells and observe real-time changes in membrane shape and protein distribution during the fusion event. This information and the tools used can then be applied to other cell biological and gene regulation questions during other types of membrane fusion.

描述（由申请人提供）： 在酵母交配过程中，核融合发生在交配对的两个单倍体核之间，形成二倍体核。因此，膜融合是受精卵形成的关键步骤，尽管对这一过程知之甚少。在最简单的模型中，内膜和外膜从纺锤体的膜半桥开始同时融合。在另一个模型中，核聚变实际上发生在三个步骤中：首先是外膜融合，然后是内膜融合，最后是纺锤体融合。为了确定核融合是否发生在一个或三个步骤中，我们正在生成CFP和YFP标记的核，ER管腔和整合膜蛋白，以遵循它们在核融合过程中转移的时间顺序。我们已经开始了我们的研究，使用核孔标记，Nic 96 p，作为核融合标记。使用Nic 96-CFP/-YFP，我们能够建立活细胞中核融合事件的时间，并观察融合事件期间膜形状和蛋白质分布的实时变化。这些信息和所使用的工具可以应用于其他类型的膜融合过程中的其他细胞生物学和基因调控问题。