CAREER: Protein degradation during the maternal-to-zygotic transition

职业：母体向合子转变过程中的蛋白质降解

• 批准号: 2046136
• 负责人: Olivia Rissland
• 金额: $ 130万
• 依托单位: University of Colorado at Denver-Downtown Campus
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-07-15 至 2026-06-30
• 项目状态: 未结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=2046136&HistoricalAwards=false
• 关键词: CAREER; Protein; degradation; during; maternal

For all life forms, development is a highly synchronized and fundamental process. The initial stages of animal development are directed by biomolecules that were made by the mother and deposited into the growing egg. Once they have served their role, these maternal biomolecules must be removed and replaced with new ones made by the embryo. This process, known as the maternal-to-zygotic transition, is essential for animal embryogenesis. Despite the importance of the maternal-to-zygotic transition, little attention has been placed on understanding how maternal proteins are removed. The goal of this CAREER award is to fill this gap through a combination of genetic and molecular biology techniques. This research will provide an entry point into three poorly understood, but critical, questions: 1) how do embryos target maternal proteins for degradation; 2) how is protein degradation controlled during development; and 3) what are the developmental consequences when this process goes awry? This award will also provide 15-month paid internships to undergraduates who identify as underrepresented minorities and study at a local undergraduate institution, Metropolitan State University of Denver (Metro). Meaningful research experiences, like the ones provided by this award, will set the students on a trajectory of success in science. This award will thus map the enormous scope of protein removal in driving embryogenesis, will transform the educational ecosystem in Denver, and ultimately will help to increase diversity in science. In Drosophila melanogaster, an E2 ubiquitin-conjugating enzyme known as “Marie Kondo” removes several maternal proteins that were critical for oogenesis but are not needed by the embryo. Marie Kondo works with an E3 ubiquitin ligase called the CTLH complex. However, little is known about how Marie Kondo and the CTLH complex are controlled. Using a combination of genetics and molecular biology, this project will investigate the mechanisms underlying the tight temporal control of substrate specificities and enzymatic activities of Marie Kondo and its partner, the CTLH complex, and determine the developmental consequences of mistiming their activity. Finally, as part of the educational program, an RNAi-based screen will be used to identify other E2s and E3s essential for the Drosophila embryogenesis, which will set the stage for future investigations in maternal protein removal during the maternal-to-zygotic transition. Together, these investigations will shed light on a new area of developmental biology—protein decay—that is critical for the creation of an adult animal from a single-cell embryo.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.

对于所有生命形式来说，发展是一个高度同步的基本过程。动物发育的最初阶段是由母亲制造的生物分子引导的，并沉积在成长的卵子中。一旦它们发挥了作用，这些母体生物分子必须被移除，并被胚胎产生的新分子取代。这一过程被称为母体到合子的转变，是动物胚胎发生所必需的。尽管母亲到合子的过渡的重要性，很少有人注意到已经放在了解母亲的蛋白质是如何被删除。这个职业奖的目标是通过遗传和分子生物学技术的结合来填补这一空白。这项研究将为三个知之甚少但至关重要的问题提供一个切入点：1）胚胎如何靶向母体蛋白质降解; 2）在发育过程中如何控制蛋白质降解; 3）当这个过程出错时，发育后果是什么？该奖项还将为那些被认定为代表性不足的少数民族的本科生提供15个月的带薪实习机会，并在当地的本科院校丹佛大都会州立大学（Metro）学习。有意义的研究经验，像这个奖项所提供的，将设置在科学成功的轨道上的学生。因此，该奖项将绘制蛋白质去除在驱动胚胎发生中的巨大范围，将改变丹佛的教育生态系统，并最终将有助于增加科学的多样性。在黑腹果蝇中，一种被称为“玛丽近藤”的E2泛素结合酶去除了几种对卵子发生至关重要但胚胎不需要的母体蛋白。玛丽近藤与E3泛素连接酶称为CTLH复合物。然而，关于近藤玛丽和CTLH复合体是如何控制的，人们知之甚少。使用遗传学和分子生物学相结合，该项目将调查的机制，严格的时间控制的底物特异性和酶活性的玛丽近藤和它的合作伙伴，CTLH复合物，并确定其活动的发展后果的时机不对。最后，作为教育计划的一部分，基于RNAi的筛选将用于鉴定果蝇胚胎发生所必需的其他E2和E3，这将为未来在母体到合子过渡期间母体蛋白去除的研究奠定基础。总之，这些研究将揭示发育生物学的一个新领域--蛋白质衰变--这对于从单细胞胚胎培育成年动物至关重要。该奖项反映了NSF的法定使命，并被认为值得通过使用基金会的智力价值和更广泛的影响审查标准进行评估来支持。