Regulation and Function of MAPKAP Kinases

MAPKAP 激酶的调控和功能

• 批准号: 6606753
• 负责人: THOMAS W STURGILL
• 金额: $ 4.08万
• 依托单位: UNIVERSITY OF VIRGINIA
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-01-01 至 2005-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6606753
• 关键词: Saccharomyces cerevisiae; alcohol phosphotransferase; biological signal transduction; chromatography; enzyme substrate complex; fungal genetics; fungal proteins; gene expression; genetic regulation; intermolecular interaction; isozymes; microarray technology; mitogen activated protein kinase; molecular genetics; molecular site; phosphoprotein phosphatase; phosphorylation; posttranslational modifications; protein engineering; protein localization; protein structure function; proteomics; recombinant proteins; site directed mutagenesis; tissue /cell culture; western blottings

DESCRIPTION (provided by applicant): This proposal is to study regulation and function of MAP kinase-activated protein kin ses (MAPKAPKs), which are important, expanding, and understudied components in the MAP kinase cascades. One key role of MAPKs is to control gene expression. MAPKs and MAPKAPKs collaborate in transcription, translation, and stabilization. Understanding this collaboration is vital to design of therapies in many diseases because MAP (mitogen-activated protein) kinase (MAPK) cascades are activated by extracellular signals (growth factors, hormones, cytokines) and intracellular signals (cellular stresses and checkpoints) as part of interconnected cascades of enzyme activations and inhibitions controlling metabolism, gene expression, and growth and development. Abnormal functions of the pathways is involved in cancer, proliferative complications of diabetes, and genetic disorders. There are three principal MAPKs (ERK, JNK, and p38 MAPK) but there are many isoforms. MAP kinases select specific targets for regulation, including members of a diverse group called collectively MAPKAPKs which extend the cascade. MAPKAPKs are related to each other by their C-terminal domains independent of whether they have one kinase domain (MNKs, MAPKAPK2) or two domains (RSKs, MSKs). Aim one is determine how MAP kinases bind and regulate their specific MAPKAP kinase targets. Aim two is to study the cellular functions of MAPKAPKs by surveying the mammalian proteome for phosphorylations inducible by MAPKAPKs. Aim three is to design and use recombinant activated COOH-terminal domains (CTDS) of RSKs and MSKs to characterize their CTD kinase activity independently of the NTD domain, and to characterize nuclear RSKs and MSKs by extraction and chromatography. Aim four is to characterize the activation, enzymatic properties, and functions of MAPKAP kinase-like enzymes (Rcklp, Rck2p) in yeast, a model eukaryote, using combined biochemical and genetic approaches.

描述（由申请人提供）：本提案旨在研究监管和 MAP 激酶激活蛋白激酶 (MAPKAPK) 的功能 MAP 激酶级联中重要的、正在扩展的、尚未被研究的成分。 MAPK 的关键作用之一是控制基因表达。 MAPK 和 MAPKAPK 在转录、翻译和稳定方面进行合作。理解 这种合作对于许多疾病的治疗设计至关重要，因为 MAP （丝裂原激活蛋白）激酶 (MAPK) 级联被激活 细胞外信号（生长因子、激素、细胞因子）和细胞内信号 信号（细胞压力和检查点）作为互连级联的一部分 控制新陈代谢、基因表达的酶激活和抑制， 以及成长和发展。通路的异常功能涉及 癌症、糖尿病的增殖性并发症和遗传性疾病。 主要有 3 种 MAPK（ERK、JNK 和 p38 MAPK），但还有很多 同工型。 MAP 激酶选择特定的调控目标，包括成员 一个称为 MAPKAPK 的多元化群体的成员，它扩展了级联。 MAPKAPK 通过其 C 端结构域相互关联，独立于 它们是否具有一个激酶结构域（MNK、MAPKAPK2）或两个结构域（RSK、 MSK）。 目标一是确定 MAP 激酶如何结合和调节其特定的 MAPKAP 激酶靶标。目标二是通过以下方式研究 MAPKAPK 的细胞功能： 调查哺乳动物蛋白质组中 MAPKAPK 诱导的磷酸化。 目标三是设计和使用重组激活的 COOH 末端结构域 (CTDS) RSK 和 MSK 独立表征其 CTD 激酶活性 NTD 结构域，并通过提取和表征核 RSK 和 MSK 色谱法。 目标四是表征激活、酶性质和功能 使用酵母（一种模型真核生物）中的 MAPKAP 激酶样酶（Rcklp、Rck2p） 结合生化和遗传学方法。