RUI: Investigating novel mechanisms of MAPKAP Kinase 2 (MK2) regulation

智力上的优点。 MAPKAP激酶2（MK2）是应激激活的p38 MAP激酶通路的重要下游靶点。该途径对于感知细胞环境中的应激是至关重要的。MK2与许多不同且重要的细胞过程相关，包括细胞周期的调节、细胞迁移和炎症的起始。MK2有两种已知的剪接变体;两者都在细胞中表达并具有催化活性。MK 2-同种型1（NM 004759.3）以17个不同的氨基酸结束，而MK 2-同种型2（NM 032960.2）以47个不同的氨基酸结束。这项研究将探讨MK2调控的新机制。特别是，该项目旨在确定通过氧化的直接修饰是否有能力调节细胞环境中的MK2，并确定亚型-1与亚型-2的调节差异。研究较多且较长的同种型-2具有充分表征的核输出（内斯）和包含在47个不同氨基酸内的核定位信号（NLS）。 这些信号传导序列使得同种型-2能够在细胞核和细胞质之间穿梭。异构体-1缺乏这些信号序列，本项目将确定异构体-1的定位。此外，亚型-2中的NLS的一部分产生了p38的结合位点，这使得亚型-2能够磷酸化和活化。 该项目将测试该结合位点的丧失如何影响亚型-1的活化特征以及亚型-1与其他蛋白质，特别是底物相互作用的能力。 这项研究将确定氧化修饰是否具有调节MK 2的能力，并将开始澄清MK 2的两种亚型在调节重要的细胞过程中是否具有不同的作用。 该项目将为本科生提供大量的研究经验，包括蛋白质的细菌表达，蛋白质纯化，哺乳动物组织培养，转染，探测蛋白质-蛋白质相互作用和激酶测定。因为肯尼索州立大学？的多样化的学生群体- 25%的少数民族和55%的女性在科学和数学学院？这个项目将有一个庞大的学生库，从其中招募少数民族和妇女到这个研究项目，并最终到干（科学，技术，工程，数学）的职业。除了在实验室的实践培训，学生将学习批判性地分析论文，并将获得科学写作和向同事做演讲的经验。在会议上和同行评审的手稿中传播结果将提供对MK2亚型的调节如何影响细胞过程（例如细胞周期，细胞迁移）的更详细的理解。该项目将加强这个主要教学机构的研究环境，为教师和学生提供必要的资源，为主要研究者和本科生提供研究经验。

Intellectual merit. MAPKAP kinase 2 (MK2) is an important downstream target of the stress activated p38 MAP kinase pathway. This pathway is critical for sensing stress in the cellular environment. MK2 has been linked to many different and important cellular processes, including regulation of the cell cycle, cellular migration, and initiation of inflammation. MK2 has two known splice variants; both are expressed and catalytically active in cells. MK2-isoform 1 (NM 004759.3) ends with 17 distinct amino acids while MK2-isoform 2 (NM 032960.2) ends with 47 distinct amino acids. This research will investigate novel mechanisms of MK2 regulation. In particular, this project aims to determine if direct modification through oxidation has the ability to regulate MK2 in the cellular environment and to determine how isoform-1 is regulated differentially from isoform-2. The more studied and longer isoform-2 has well characterized nuclear export (NES) and nuclear localization signals (NLS) contained within the 47 distinct amino acids. These signaling sequences enable shuttling of isoform-2 between the nucleus and cytoplasm. Isoform-1 lacks these signal sequences, and this project will determine the localization of isoform-1. Additionally, a portion of the NLS in isoform-2 creates a binding site for p38, which enables phosphorylation and activation of isoform-2. This project will test how loss of that binding site impacts both the activation profile of isoform-1 and the ability of isoform-1 to interact with other proteins, particularly substrates. This research will identify if oxidative modification has the ability to regulate MK2 and will begin to clarify if the two isoforms of MK2 have different roles in regulating important cellular processes.Broader impacts. This project will provide undergraduate students with a multitude of research experiences, including bacterial expression of proteins, protein purification, mammalian tissue culture, transfection, probing protein-protein interactions and kinase assays. Due to Kennesaw State University?s diverse student population - 25% minority and 55% female in the College of Science and Mathematics ? this project will have a large pool of students from which to recruit minorities and women to this research project and ultimately to STEM (Science, Technology, Engineering, Mathematics) careers. In addition to hands-on training in the lab, students will learn to critically analyze papers and will gain experience in scientific writing and making presentations to colleagues. Dissemination of the results at meetings and in peer reviewed manuscripts will provide a more detailed understanding of how the regulation of MK2 isoforms may impact cellular processes (e.g. the cell cycle, cellular migration). This project will strengthen the research environment at this predominantly teaching institution for faculty and students alike, providing essential resources for the Principal Investigator and research experiences for undergraduate students.