GAMETE MEMBRANE ADHESINO AND FUSION DURING FERTILIZATION

受精过程中配子膜的粘附和融合

• 批准号: 7919159
• 负责人: William J Snell
• 金额: $ 10.91万
• 依托单位: UT SOUTHWESTERN MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-09-30 至 2012-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7919159
• 关键词: Actins; Adhesions; African Trypanosomiasis; Algae; Animals; Binding; Biochemical; Biological; Cell fusion; Cell membrane; Cell surface; Cells; Chimeric Proteins; Chlamydomonas; Cnidaria; Deletion Mutation; Disease; Event; Family member; Female; Fertilization; Funding; Future; Gene Targeting; Genes; Genetic; Germ Cells; Goals; Human; Insecta; Insertional Mutagenesis; Laboratories; Malaria; Membrane; Membrane Fusion; Methods; Modeling; Molecular; Mutation Analysis; Organism; Parasites; Partner in relationship; Plasmodium; Plasmodium berghei; Positioning Attribute; Property; Proteins; Protozoa; Reproduction; Rodent; Role; Signal Pathway; Signal Transduction; Site; Sperm-Ovum Interactions; Structure; Surface; System; Testing; Vascular Plant; deletion analysis; human disease; insight; male; protein function; sex; transmission process

The long-term goals of my laboratory are to understand the cellular and molecular mechanisms that underlie gamete interactions and cell-cell fusion during fertilization. We use the unicellular, biflagellated alga Chlamydomonas for our studies, an organism whose fertilization is highly amenable to cell biological, biochemical, molecular biological, and genetic approaches. Importantly, during Chlamydomonas fertilization, the interacting male and female gametes undergo the cell biological events that characterize fertilization in almost all animals. Understanding gamete fusion is needed to inform future studies on human reproduction. In addition, gamete fusion is essential for insect transmission of several devastating human diseases caused by parasitic protozoa, including the organisms that cause malaria and sleeping sickness. Knowing the molecular mechanisms of gamete fusion in parasitic protozoa would be an important advance in efforts to reduce the impact of the diseases they cause. Unfortunately, in spite of the many different fertilization systems being studied, our molecular understanding of gamete fusion in any organism remains rudimentary. The objectives of this proposal are to characterize the cellular and molecular mechanisms of gamete fusion in Chlamydomonas, focusing on two gamete fusion proteins, Fus1 a female gamete-specific protein, and a new male gamete-specific protein we discovered in this funding period, FusM. Both are localized at the sites of fusion and essential for fusion. Unlike all other gamete fusion proteins identified to date in any organism, FusM is widely conserved. Although not found in bilaterian animals, FusM family members are present in simple animals, higher plants, and pathogenic protozoa. In collaborative efforts using targeted gene disruption in Plasmodium berghei, a rodent malaria parasite, we discovered that the Plasmodium FusM is essential for gamete fusion. Thus, our studies on Chlamydomonas have revealed a conserved mechanism for gamete fusion. Now that we have a protein on each Chlamydomonas gamete that is essential for gamete fusion, we are in a unique position to make new insights into fundamental cellular and molecular mechanisms of gamete fusion. We propose to identify the binding partners of Fus1 and FusM, to determine the molecular functions of the proteins in gamete membrane adhesion and fusion, to investigate the molecular mechanisms of their localization, and to characterize their molecular properties before, during, and after fusion.

我的实验室的长期目标是了解构成其基础的细胞和分子机制 受精过程中配子的相互作用和细胞-细胞融合。我们用单细胞、双鞭毛的藻类 衣藻在我们的研究中，是一种受精高度依赖细胞生物学的有机体， 生物化学、分子生物学和遗传学方法。重要的是，在衣藻受精期间， 相互作用的雄配子和雌配子经历细胞生物学事件，这些事件是受精的特征 几乎所有的动物。了解配子融合是未来人类生殖研究的基础。 此外，配子融合对于昆虫传播几种毁灭性的人类疾病是必不可少的。 由寄生原生动物，包括引起疟疾和昏睡病的有机体。了解 寄生原生动物配子融合的分子机制将是研究 减少它们引起的疾病的影响。不幸的是，尽管有许多不同的受精方式 在系统的研究中，我们对任何生物体中配子融合的分子理解仍然处于初级阶段。 这项建议的目的是描述配子融合的细胞和分子机制。 在衣藻中，集中在两个配子融合蛋白上，FUS1是一种雌配子特有的蛋白，而一个是 我们在这个资助期发现了新的雄配子特异蛋白FusM。这两个网站都是本地化的 是聚变的关键，也是聚变的关键。与迄今为止在任何生物体中发现的所有其他配子融合蛋白不同， FusM是广泛保守的。虽然在双边动物中没有发现，但FusM家族成员存在于 简单的动物、高等植物和致病原生动物。使用靶向基因的协作努力 伯氏疟原虫是一种啮齿动物疟疾寄生虫，我们发现该疟原虫是 配子融合所必需的。因此，我们对衣藻的研究揭示了一种保守的机制 用于配子融合。现在我们在每个衣藻配子上都有一种对配子必不可少的蛋白质 融合，我们处于一个独特的位置，可以对基本的细胞和分子做出新的见解 配子融合的机制。我们建议确定FUS1和FusM的结合伙伴，以确定 研究这些蛋白质在配子膜黏附和融合中的分子功能。 其定位的分子机制，并表征其分子性质之前，期间，和 在融合之后。