Genetic Investigation of Lammer Protein Kinase Structure and Function in Drosophila

果蝇 Lammer 蛋白激酶结构和功能的遗传研究

• 批准号: 9724006
• 负责人: Bruce Chase
• 金额: $ 28.95万
• 依托单位: University of Nebraska Medical Center
• 依托单位国家: 美国
• 项目类别: Continuing grant
• 财政年份: 1997
• 资助国家: 美国
• 起止时间: 1997-09-01 至 2001-08-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=9724006&HistoricalAwards=false
• 关键词: Genetic; Investigation; Lammer; Protein; Kinase

ABSTRACT 9724006 RABINOW Dr. Rabinow's laboratory is characterizing a Drosophila gene known as Doa, which encodes a member of the LAMMER kinases, a recently discovered eukaryotic family of protein kinases. Protein kinases transfer phosphate groups to other proteins as a means of regulating their activity, and regulate many essential cellular processes. Homologues to Doa throughout the animal kingdom have very similar amino-acid sequences in the part of the protein that has enzymatic activity, suggesting that they have equally similar function in cellular processes. Recessive Doa mutations in flies are almost lethal; however, rare adults do survive, but show defects in eye development. Other defects in Doa mutants include failure of a normal embryonic central nervous system (CNS) to develop and segmentation defects. The DOA kinase is widely expressed and is found at particularly high levels in the embryonic CNS, where rapid cell division and differentiation occurs. The DOA kinase comes in two major forms that differ in size. The larger of these (105 kD) is found only in the cell cytoplasm, whereas the smaller version (55 kD) is also found in the cell nucleus. The structure of the 105 kD cytoplasmic form is completely unknown. In the current research project, Dr. Rabinow's laboratory will carry out further structural and functional characterization of DOA kinase to define the role of this kinase family in the initiation and maintenance of cellular differentiation. The first task will be to determine the structure of the 105 kD protein. In addition, antibodies against the two protein isoforms will be generated to examine their expression. Drosophila expressing specifically altered forms of the kinase will be constructed to determine whether the two protein isoforms perform different functions, as well as to provide more complete genetic characterization of Doa mutant defects. These data will be used to identify one or more of the cellular processes that are altered in mutants. R esults from this project will substantially advance the long-term goal of elucidating the specific cellular functions of the LAMMER protein kinase family and the developmental signaling pathway(s) in which they function.

摘要9724006拉比诺博士的实验室正在研究一种名为doA的果蝇基因，该基因编码最近发现的真核蛋白激酶家族Lammer蛋白激酶的成员之一。蛋白激酶将磷酸基团转移到其他蛋白质上，作为一种调节其活性的手段，并调节许多重要的细胞过程。在整个动物界，DOA的同源物在具有酶活性的蛋白质部分具有非常相似的氨基酸序列，这表明它们在细胞过程中具有同样相似的功能。果蝇的隐性DOA突变几乎是致命的；然而，很少有成虫存活下来，但在眼睛发育方面显示出缺陷。DoA突变体的其他缺陷包括正常胚胎中枢神经系统(CNS)发育失败和分段缺陷。DOA激酶在胚胎中枢神经系统中广泛表达，并且水平特别高，在那里细胞发生快速分裂和分化。DOA激酶有两种不同大小的主要形式。其中较大的(105kD)仅在细胞质中发现，而较小的(55kD)也在细胞核中发现。105kD细胞质形式的结构完全未知。在目前的研究项目中，Rabinow博士的实验室将对DOA激酶进行进一步的结构和功能表征，以确定该激酶家族在启动和维持细胞分化中的作用。第一项任务是确定105kD蛋白质的结构。此外，还将产生针对这两种蛋白质亚型的抗体来检测它们的表达。将构建表达特定改变形式的激酶的果蝇，以确定这两种蛋白异构体是否发挥不同的功能，以及提供更完整的DOA突变缺陷的基因特征。这些数据将被用来识别在突变中改变的一个或多个细胞过程。该项目的结果将极大地推进阐明Lammer蛋白激酶家族的特定细胞功能以及它们在其中发挥作用的发育信号通路(S)的长期目标。