Investigation of the function and regulation of ERK3

描述(由申请人提供)： MAPK与相关的细胞周期蛋白依赖性蛋白激酶(CDKs)一样，是细胞对有丝分裂刺激、发育和环境应激反应的重要组成部分。最近，越来越多的证据表明，ERK3是MAPK家族的成员之一，在包括烟草相关鳞状细胞癌在内的许多不同类型的癌症中可能被解除调控。然而，研究ERK3的基础生物学的研究相对较少。因此，不仅从基础科学的角度对ERK3进行进一步的研究，而且作为癌症治疗的潜在治疗或诊断靶点也是有必要的。最近，我们发现ERK3的亚细胞定位在细胞周期中是有时间调节的。因此，我们认为，进一步研究ERK3的功能可能会对哺乳动物细胞周期的控制以及最终对癌症的控制提供更多的了解。这一建议将探讨(1)ERK3在高尔基体生物学中的作用，(2)在细胞周期的调节中，以及(3)有丝分裂激酶PLK和cdc2/Cyclin B1是否在体内调节ERK3。SiRNA技术结合共聚焦显微镜将被用来探索ERK3在高尔基体中的作用。这些实验将专门研究ERK3是否参与有丝分裂过程中高尔基体碎裂的调节，或者是正常细胞周期进程所必需的。为了研究PLK或cdc2/Cyclin B1是否在体内调节ERK3，我们将使用生化和细胞生物学相结合的方法来研究这种关联的时机。在这些研究之后，将进行体外生化筛选，以确定PLK和/或cdc2/Cyclin B1靶向的ERK3的特定残基或区域。这些结构域在体内与ERK3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.