Investigation of the function and regulation of ERK3

描述（由申请人提供）： MAPK和相关的细胞周期蛋白依赖性激酶（cdks）一样，是细胞对促有丝分裂刺激、发育和环境应激反应的非常重要的部分。最近，越来越多的证据表明，ERK 3，MAPK家族的一员，可能会在许多不同类型的癌症，包括烟草相关的鳞状细胞癌失调。然而，对ERK 3的基础生物学研究相对较少。因此，对ERK 3的进一步研究不仅从基础科学的角度来看是必要的，而且作为癌症治疗中的潜在治疗或诊断靶点也是必要的。最近，我们发现ERK 3的亚细胞定位在细胞周期中受到时间调节。因此，我们建议，进一步调查的ERK 3的功能可能会提供一个额外的水平的了解控制的哺乳动物细胞周期，并最终癌症。本研究将探讨ERK 3在高尔基体生物学中的作用，ERK 3在细胞周期调控中的作用，以及ERK 3在体内是否受有丝分裂激酶PLK和cdc 2/cyclin B1的调控。SiRNA技术结合共聚焦显微镜将用于探索ERK 3在高尔基体中的作用。这些实验将具体解决ERK 3是否参与有丝分裂期间高尔基体片段化的调节或正常细胞周期进程所需。要调查是否PLK或cdc 2/细胞周期蛋白B1调节ERK 3在体内，我们将调查这种关联的时间，通过使用相结合的生物化学和细胞生物学方法。这些研究之后将进行体外生物化学筛选，旨在鉴定PLK和/或cdc 2/细胞周期蛋白B1靶向的ERK 3的特定残基或区域。这些结构域与体内ERK 3功能的生物学相关性将通过评估其亚细胞定位和对细胞周期的影响来评估。

DESCRIPTION (provided by applicant): MAPKs like the related cyclin dependent kinases (cdks) are a very important part of the cellular response to mitogenic stimuli, development and environmental stresses. Recently, there has been increasing evidence that ERK3, a member of the MAPK family, may be deregulated in a number of different types of cancer including tobacco associated squamous cell carcinomas. There have been, however, relatively few studies investigating the basic biology of ERK3. Thus, further investigation of ERK3 is warranted not only from a basic science point of view, but also as a potential therapeutic or diagnostic target in the treatment of cancer. Recently, we have discovered that the subcellular localization of ERK3 is temporally regulated during the cell cycle. We therefore propose that further investigation of the function of ERK3 may provide an additional level of understanding of the control of the mammalian cell cycle and ultimately of cancer. This proposal will explore (1) The role of ERK3 in the biology of the Golgi, (2) In the regulation of the cell cycle, and (3) Whether the mitotic kinases PLK and cdc2/cyclin B1 regulate ERK3 in vivo. SiRNA technology combined with confocal microscopy will be used to explore the role of ERK3 in the Golgi. These experiments will specifically address whether ERK3 is involved in the regulation of the fragmentation of the Golgi during mitosis or is required for normal cell cycle progression. To investigate whether PLK or cdc2/cyclin B1 regulate ERK3 in vivo, we will investigate the timing of this association by using a combined biochemical and cell biological approach. These studies will be followed by an in vitro biochemical screen aimed to identify the specific residues or regions of ERK3 targeted by PLK and/or cdc2/cyclin B1. The biological relevance of these domains to ERK3 function in vivo will be assessed by an evaluation of its subcellular localization and effect on the cell cycle.