REGULATION OF SKIN HOMEOSTASIS BY RNA-BINDING PROTEINS

RNA结合蛋白对皮肤动态平衡的调节 项目摘要/摘要 每四个美国人中就有一个受到皮肤病的影响，这强烈地推动了人们对皮肤病机制的理解 潜在的表皮动态平衡。RNA结合蛋白(RBPs)与主要的精氨酸甲基转移酶 PRMT1是控制角质形成细胞增殖和分化所必需的。我们已经确认了两个 核仁RNA解旋酶和多个PRMT1靶点是正常皮肤动态平衡所必需的。我们有 还确定PRMT1本身是RBP。我研究的一个长期目标是了解动态平衡 皮肤中的作用机制；本K01应用程序的目标是表征RBPs在表皮中的作用 动态平衡，特别是两个解旋酶，PRMT1和PRMT1靶向限制性商业惯例。 在目的I中，我们将测试我们的模型，PRMT1‘S维持角质形成细胞的祖细胞状态。我们的 初步数据表明，PRMT1-的五个稳定的RBP相互作用元件也是ITS的靶标 甲基转移酶活性--也是维持体内平衡所必需的。在Aim IA中，我们测试了 这些限制性商业惯例上的精氨酸甲基化。在Aim IB中，我们测试了这些限制性商业惯例及其甲基化对基因的影响 在角质形成细胞中的表达。 在AIM II中，我们测试了RNA解旋酶如何促进角质形成细胞分化的模型。我们的数据显示 两种解旋酶是序列特异性的前mRNA结合蛋白，通过离开 核仁调节剪接和潜在的转录。我们的结果还表明，这些解旋酶可以 形成同二聚体和杂二聚体，其中杂二聚体是相互作用最强的。在AIM IIA中，我们将测试他们的 在角质形成细胞分化的转录控制中的作用及其对剪接的明显控制是否是直接的。 在AIM IIB中，我们将评估它们的二聚化状态及其与它们在动态平衡、剪接和 抄写。 我的背景是RBP生物化学、生物信息学、方法开发和CLIP-SEQ。在整个过程中 在斯坦福大学皮肤科的环境下，我计划获得 精通(1)上皮生物学、(2)蛋白质组学、(3)机器学习和(4)统计学。我的研究兴趣是 皮肤生物学需要这种广泛的技能，而这个研究计划将为我独立的职业生涯做好准备 致力于研究上皮组织中基因调控机制的学术科学家。

REGULATION OF SKIN HOMEOSTASIS BY RNA-BINDING PROTEINS PROJECT SUMMARY/ABSTRACT One in four Americans are affected by skin disease, intensely motivating an understanding on the mechanisms underlying epidermal homeostasis. RNA-binding proteins (RBPs) and the main arginine methyltransferase PRMT1 are required for the control of keratinocyte proliferation and differentiation. We have identified two nucleolar RNA helicases and multiple PRMT1 targets that are required for normal skin homeostasis. We have also identified PRMT1 itself is an RBP. A long-term goal of my research is to understand homeostatic mechanisms in skin; the goal of this K01 application is to characterize the roles of RBPs in epidermal homeostasis, specifically the two helicases, PRMT1 and PRMT1 target RBPs. In Aim I, we will test our model for PRMT1’s maintenance of the progenitor state in keratinocytes. Our preliminary data indicates that five stable RBP interactors of PRMT1 – which are also targets of its methyltransferase activity – are also required to maintain homeostasis. In Aim IA, we test the consequence of arginine methylation on these RBPs. In Aim IB, we test the effect of these RBPs and their methylation on gene expression in keratinocytes. In Aim II, we test our model for how RNA helicases promote keratinocyte differentiation. Our data suggests two helicases are sequence-specific, pre-mRNA-binding proteins that promote differentiation by departing the nucleolus to regulate splicing and potentially transcription. Our results also suggest that these helicases can form homo- and heterodimers, with the heterodimer being the strongest interaction. In Aim IIA, we will test their role in transcriptional control of keratinocyte differentiation and whether their apparent control of splicing is direct. In Aim IIB, we will evaluate their dimerization state and its relation to their role in homeostasis, splicing and transcription. My background is in RBP biochemistry, bioinformatics, methods development, and CLIP-seq. Over the course of the training period and within the environment of Stanford’s Department of Dermatology, I plan to acquire expertise in (1) epithelial biology, (2) proteomics, (3) machine learning, and (4) statistics. My research interest in skin biology requires this broad skill set, and this research plan will prepare me for an independent career as an academic scientist working on the mechanisms of gene regulation in epithelial tissue.