Molecular regulation of germline development

项目总结/摘要 我们的目标是了解生殖细胞调控的分子原理。我们专注于一个决定， 细胞使：自我更新或开始分化。这一决定在所有组织中都是基本的， 人体健康促进干细胞状态的调节器必须活跃以驱动自我更新和全能性， 但必须失活分化;相反，对分化至关重要的调节因子必须是 在干细胞中是不活跃的，但在正确的时间和分化的地方被激活。调控 从自我更新到分化的过渡平衡了两种状态，是发展的根本， 体内平衡和人类健康。这种平衡的缺陷会导致人类疾病，包括组织萎缩和 癌 RNA调控是从蠕虫到哺乳动物生殖细胞中的一个主要保守主题。关键RNA结合蛋白 调节动物生殖过程中的自我更新和分化。我们专注于PUF（Pumilio和FBF）RNA- 结合蛋白，在动物物种的生殖细胞和神经生物学中具有关键作用。在人类中， 有缺陷的PUF蛋白导致不育和癫痫。PUF蛋白有许多共同的特征： 转录组和蛋白质组通过结合超过1000种mRNA，其中100种保守， 线虫和人类;它们通过多种机制调节靶mRNA，包括激活和 抑制;并且它们以组合的方式与伴侣蛋白一起工作，其中大多数也是保守的。 现在的挑战是了解PUF蛋白如何调节自我更新和自我更新之间的平衡。 在分子方面和在天然的体内环境中的分化。我们的建议通过以下方式应对这一挑战： 聚焦四个问题。首先，我们阐明了不同的PUF机制-激活和抑制-工作 共同控制生殖细胞的命运其次，我们阐明了PUF伙伴关系如何在体内运作，以了解 他们的逻辑和收集一般原则。第三，我们阐明了自我更新与 分化第四，我们阐明了多个PUF蛋白和伙伴的功能模块如何维持 干细胞在不同的生理和环境条件下。这四个问题对所有人来说都是根本性的 PUF法规。对于每一个，我们采取多学科和多尺度的方法，并利用一个特殊的 易处理的网络总之，我们的发现将促进我们对生殖细胞调控的理解， 广泛的影响，包括人类的新疗法设计。

PROJECT SUMMARY/ABSTRACT Our goal is to understand molecular principles of germ cell regulation. We focus on a decision that all stem cells make: to self-renew or begin differentiation. This decision is fundamental in all tissues and is critical to human health. Regulators that promote the stem cell state must be active to drive self-renewal and totipotency, but must be inactivated for differentiation; conversely, regulators that are critical for differentiation must be inactive in stem cells, but then become activated at the right time and place for differentiation. Regulation of the transition – from self-renewal to differentiation – balances the two states and is fundamental to development, homeostasis and human health. Defects in that balance cause human disease, including tissue atrophy and cancer. RNA regulation is a major conserved theme in germ cells from worms to mammals. Key RNA-binding proteins regulate self-renewal and differentiation across animal phylogeny. We focus on PUF (Pumilio and FBF) RNA- binding proteins, which have pivotal roles in germ cells and neurobiology across animal species. In humans, defective PUF proteins cause infertility and epilepsy. PUF proteins have many features in common: They sculpt the transcriptome and proteome by binding to more than 1000 mRNAs, with 100's conserved between nematodes and humans; they regulate target mRNAs by diverse mechanisms, including activation and repression; and they work in a combinatorial fashion with partner proteins, most of which are also conserved. The challenge now is to understand how PUF proteins regulate the balance between self-renewal and differentiation in molecular terms and in a native in vivo context. Our proposal addresses this challenge with a sharp focus on four issues. First, we elucidate how distinct PUF mechanisms – activation and repression – work together to control germ cell fates. Second, we elucidate how PUF partnerships operate in vivo, to understand their logic and glean general principles. Third, we elucidate regulation of the balance between self-renewal to differentiation. Fourth, we elucidate how a functional module of multiple PUF proteins and partners maintains stem cells under diverse physiological and environmental conditions. These four issues are fundamental to all PUF regulation. For each, we take a multidisciplinary and multiscale approach and exploit an exceptionally tractable network. Together, our findings will advance our understanding germ cell regulation with potential for broad impact, including of new therapy designs for humans.