Molecular regulation of germline development

项目概要/摘要 我们的目标是了解生殖细胞调节的分子原理。我们专注于一切源于的决定 细胞进行：自我更新或开始分化。这一决定对于所有组织都是基础性的，并且对于 人类健康。促进干细胞状态的调节因子必须活跃以驱动自我更新和全能性， 但必须失活以进行分化；相反，对于差异化至关重要的监管机构必须 在干细胞中不活跃，但随后在正确的时间和地点被激活以进行分化。监管 从自我更新到分化的转变平衡了两种状态，是发展的基础， 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 癌症。 RNA 调控是从蠕虫到哺乳动物的生殖细胞中的一个主要保守主题。关键 RNA 结合蛋白 调节动物系统发育的自我更新和分化。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发挥着关键作用。在人类中， 有缺陷的 PUF 蛋白会导致不孕和癫痫。 PUF 蛋白有许多共同特征： 转录组和蛋白质组通过与 1000 多个 mRNA 结合，其中 100 个 mRNA 是保守的 线虫和人类；它们通过不同的机制调节目标 mRNA，包括激活和 镇压；它们与伴侣蛋白以组合方式发挥作用，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 分子术语和天然体内环境中的差异。我们的建议解决了这一挑战 重点突出四个问题。首先，我们阐明不同的 PUF 机制（激活和抑制）如何发挥作用 共同控制生殖细胞的命运。其次，我们阐明 PUF 伙伴关系如何在体内运作，以了解 他们的逻辑和收集的一般原则。第三，我们阐明了自我更新与自我更新之间的平衡调节。 差异化。第四，我们阐明了多个 PUF 蛋白和伙伴的功能模块如何维持 不同生理和环境条件下的干细胞。这四个问题对于所有人来说都是至关重要的 PUF调节。对于每一个，我们都采取多学科和多尺度的方法，并利用特殊的方法 易于处理的网络。总之，我们的研究结果将促进我们对生殖细胞调控的理解，并具有潜在的潜力 广泛的影响，包括针对人类的新疗法设计。

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.