REGULATION OF SKIN HOMEOSTASIS BY RNA-BINDING PROTEINS

RNA结合蛋白对皮肤稳态的调节 项目总结/摘要 四分之一的美国人受到皮肤病的影响，强烈激发对机制的理解 潜在的表皮稳态。RNA结合蛋白（RBP）和主要的精氨酸甲基转移酶 PRMT 1是控制角质形成细胞增殖和分化所必需的。我们已经确定了两个 核仁RNA解旋酶和多个PRMT 1靶点是正常皮肤稳态所需的。我们有 还鉴定了PRMT 1本身是RBP。我研究的一个长期目标是了解 皮肤中的机制;该K 01应用程序的目标是表征RBP在表皮中的作用 在一些实施方案中，两种解旋酶PRMT 1和PRMT 1靶向RBP。 在目的I中，我们将测试我们的模型PRMT 1在角质形成细胞中的祖细胞状态的维持。我们 初步数据表明，PRMT 1的五个稳定的RBP相互作用物-这也是其靶点， 甲基转移酶活性-也需要维持体内平衡。在Aim IA中，我们测试以下结果： 精氨酸的甲基化。在目的IB中，我们测试了这些RBP及其甲基化对基因表达的影响。 在角质形成细胞中的表达。 在目的II中，我们测试我们的模型如何RNA解旋酶促进角质形成细胞分化。我们的数据表明 两种解旋酶是序列特异性的前mRNA结合蛋白，其通过使前mRNA结合蛋白与前mRNA结合蛋白分离而促进分化。 核仁来调节剪接和潜在的转录。我们的研究结果还表明，这些解旋酶可以 形成同二聚体和异二聚体，其中异二聚体是最强的相互作用。在Aim IIA中，我们将测试 在角质形成细胞分化的转录控制中的作用以及它们对剪接的表观控制是否是直接的。 在Aim IIB中，我们将评估它们的二聚化状态及其与它们在稳态、剪接和 转录。 我的背景是RBP生物化学，生物信息学，方法开发和CLIP-seq。过程中 在培训期间和斯坦福大学皮肤科的环境中，我计划获得 专业知识（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，这个研究计划将为我作为一个独立的职业生涯做好准备。 研究上皮组织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.