RUI: RSK & Rock Phosphorylation of NHE1 Regulates Cell Motility Events in CCL39 Fibroblasts

钠氢交换器(NHE)是一种膜结合蛋白，参与调节许多重要的细胞功能。细胞如何运动是由各种不同的蛋白质同步的一系列复杂的事件。Nhe是指导细胞运动的关键蛋白质之一，因为交换器起到极化细胞并协调细胞骨架的作用。NHE是如何控制细胞运动的，人们对此只有部分了解。本项目将研究核糖体S-6激酶(RSK)和RhoA激酶(ROCK)对NHE细胞内尾部的磷酸化作用。该项目的假设是，RSK和ROCK都能调节NHE在细胞运动中的作用。该项目将首先确定ROCK结合和磷酸化NHE的位置，然后确定这些磷酸化位点如何协调NHE的影响，包括细胞骨架组织、定向细胞运动和蛋白质-蛋白质相互作用。NHE是被认为参与控制细胞极性、细胞骨架组织和多种细胞类型中信号复合体形成的几种转运蛋白之一。这个项目的结果将有助于我们理解一个重要的生长模型，解释细胞骨架调节和运动的细胞生物学。更广泛的影响：这个项目的一个重要部分是为本科生提供现实和有意义的研究。这个项目的目的是让学生完全沉浸在研究体验中，并让学生充分受益于将研究纳入大学的课程内外。研究人员在培养本科生研究人员方面有着长期的承诺和卓越的记录。他们实验室的校友已经进入了加州大学圣地亚哥分校、麻省理工学院、耶鲁大学、明尼苏达大学、爱荷华州大学和许多工业岗位的研究生院。这个项目将涉及生物化学和细胞生物学研究的重要方面的本科生，这项工作将加深对运输蛋白的理解，因为这些膜蛋白有助于调节细胞迁移事件。之前资助的瑞奖为21名本科生和8名高中生提供了研究机会。在这29名学生中，19名是女性，3名是美洲原住民，2名是西班牙裔。资助这一研究项目将支持S研究员提供训练有素的本科生进入研究生院、职业学校和劳动力市场的能力。具体地说，该项目将为另外12至15名本科生和2至3名高中生提供课外研究经验，相当于24至30年的学术研究经验。此外，另有18至24名学生将获得一年的研究经验，作为生物化学和生物技术专业为期一年、10学分的高级课程的一部分。最后，小组中的9名学生将有机会参加全日制暑期研究体验。

The sodium hydrogen exchanger (NHE) is a membrane bound protein involved in regulating many important cellular functions. How a cell moves is a complex series of events synchronized by a variety of different proteins. NHE is one of the proteins critical in directing cell movement as the exchanger acts to polarize the cell and coordinates the cytoskeleton. How NHE controls cell motility is only partially understood. This project will investigate the phosphorylation of the intracellular tail of NHE by two protein kinases, Ribosomal S-6 Kinase (RSK) and RhoA Kinase (ROCK). The hypothesis of this project is that both RSK and ROCK function to regulate NHE's role in cell movement. This project will first determine where ROCK binds and phosphorylates NHE and then identify how each of these phosphorylation sites coordinates the effects of NHE including cytoskeletal organization, directed cell motility, and protein-protein interactions. NHE is one of several transport proteins thought to be involved in controlling cell polarity, cytoskeletal organization and the formation of signaling complexes in a wide variety of cell types. The results of this project will contribute our understanding of an important growing model explaining the cell biology of cytoskeletal regulation and motility. Broader Impacts: A significant portion of this project is to provide realistic and meaningful research for undergraduates. This project is designed to fully immerse students into a research experience and allow students to benefit fully from the incorporation of research both within the curriculum and outside the curriculum at the university. The investigators have a long track record of commitment and excellence in training undergraduate researchers. Alumni from their labs have gone onto graduate school at UCSD, MIT, Yale, U of MN, Iowa U and many industrial positions. This project will involve undergraduates in important aspects of biochemistry and cellular biology research and together, this work will further the understanding of transport proteins as these membrane proteins aid in regulating cell migration events. A previously funded RUI Award provided research opportunities for 21 undergraduates and 8 high school students. Of these 29 students, 19 were women, 3 were Native Americans, and 2 were Hispanic. Funding this research project will sustain the investigator?s ability to provide well-trained undergraduates to enter graduate school, professional school, and the workforce. Specifically this project will provide another 12 to 15 undergraduates and 2 to 3 high school students research experiences outside the curriculum that will equate to 24 to 30 academic years of research experience. In addition, another 18 to 24 students will gain one year of research experience each as part of a year-long, 10 credit senior capstone course for the Biochemistry and Biotechnology Major. Finally, 9 students in the group will be provided the opportunity to participate in a full time summer research experience.