NOVEL KINASE-HOMOLOGOUS NUCLEAR PROTEIN

新型激酶同源核蛋白

基本信息

批准号：
3568434
负责人：
Kristen M Johansen
金额：
$ 4.75万
依托单位：
IOWA STATE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
1994
资助国家：
美国
起止时间：
1994-04-01 至 1996-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/3568434
关键词：
Drosophilidae SDS polyacrylamide gel electrophoresis biological signal transduction cell cycle chimeric proteins chromatin complementary DNA embryogenic cleavage enzyme activity epitope mapping gene mutation genetic mapping immunocytochemistry laboratory mouse laboratory rabbit molecular cloning monoclonal antibody nuclear matrix nucleoproteins protein kinase protein structure function protooncogene serine proteinases site directed mutagenesis western blottings

Drosophilidae SDS polyacrylamide gel electrophoresis biological signal transduction cell cycle chimeric proteins chromatin complementary DNA embryogenic cleavage enzyme activity epitope mapping gene mutation genetic mapping immunocytochemistry laboratory mouse laboratory rabbit molecular cloning monoclonal antibody nuclear matrix nucleoproteins protein kinase protein structure function protooncogene serine proteinases site directed mutagenesis western blottings

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项目摘要

One of the major foci in cell biology is to understand the process of nuclear division. In each cell cycle the chromosomes must be faithfully replicated and the complex nuclear structure has to be duplicated and reorganized. Intensive research has been directed towards identifying the signal transduction pathways and molecules involved in the complex and dynamic events required for mitosis and cell division. This research has demonstrated that many of the molecules playing important roles in control of the cell cycle are protein kinases. Of particular medical significance is the finding that many cancers appear to stem from alterations in such cell cycle regulatory molecules which lead to genomic instability and tumorigenesis, and these molecules are highly evolutionary conserved. Thus the study of mitosis and cell division in various species promises to help elucidate general principles governing cell division which apply to humans as well. The object of this proposal is to clone and analyze the molecular structure and functional biology of a novel nuclear protein (p2Ab17) in Drosophila melanogaster which shows a dynamic cell cycle-specific distribution pattern and shares homology with serine-threonine kinases related to the raf family of oncogenes. This protein was first identified immunocytochemcally using monoclonal antibody 2A. The proposed experiments are designed to elucidate structural and biochemical features of the p2Ab17 protein, which co-localizes with the mitotic apparatus and chromosomes at metaphase and with chromatin or the nuclear matrix at interphase, and to determine the role p2Ab17 may play in signal transduction processes involved in nuclear division. Drosophila embryos provide an especially advantageous model system for these studies due to their synchronous early syncytial cleavage pattern which can be readily investigated using molecular, cell biological, and genetic approaches. We will apply these techniques in order to initiate a study of regulatory controls for nuclear division and to determine the functional role of p2Abl7 during mitosis. Since cancer could be viewed as a disease of the signaling system that controls cell proliferation and differentiation, understanding the roles of such signaling molecules in mitosis during normal Drosophila development will yield important insight into how they may function in both normal and cancerous cells in humans.

细胞生物学的主要焦点之一是了解核部门。在每个细胞周期中，染色体必须忠实复制，复杂的核结构必须重复，并且重组。密集研究是针对确定的涉及复合物的信号转导途径和分子有丝分裂和细胞分裂所需的动态事件。这项研究有证明许多分子在控制中起重要作用细胞周期的是蛋白激酶。特殊的医学意义是许多癌症似乎源于这种改变的发现细胞周期调节分子，导致基因组不稳定性和肿瘤发生，这些分子是高度进化的。因此各种物种中有丝分裂和细胞分裂的研究有望帮助阐明适用于人类的细胞分区的一般原则也是如此。该建议的目的是克隆和分析分子新型核蛋白的结构和功能生物学（P2AB17）果蝇Melanogaster显示了动态细胞周期特异性的分布模式并与丝氨酸 - 硫代激酶共享同源性与RAF的癌基因家族有关。该蛋白首先被鉴定出来使用单克隆抗体2a进行免疫细胞化学。提出的实验旨在阐明结构和生化特征 P2AB17蛋白，与有丝分裂设备共定位，中期和染色质或核基质处的染色体相间，并确定p2ab17的作用可能在信号中扮演核部涉及的转导过程。果蝇胚胎为这些研究提供了特别有利的模型系统他们同步的早期合成裂解模式可以很容易地使用分子，细胞生物学和遗传方法进行了研究。我们将应用这些技术以启动监管研究控制核分裂并确定的功能作用 P2ABL7在有丝分裂过程中。由于癌症可以看作是信号系统的疾病控制细胞增殖和分化，了解角色正常果蝇期间有丝分裂中这种信号分子的开发将对它们如何发挥作用产生重要的见解人类的正常细胞和癌细胞。