Chitinase-related Growth Factors in Drosophila

果蝇几丁质酶相关生长因子

• 批准号: 9728197
• 负责人: Peter Bryant
• 金额: $ 30万
• 依托单位: University of California-Irvine
• 依托单位国家: 美国
• 项目类别: Continuing Grant
• 财政年份: 1998
• 资助国家: 美国
• 起止时间: 1998-01-01 至 2001-12-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=9728197&HistoricalAwards=false
• 关键词: Chitinase; related; Growth; Factors; Drosophila

Peter J. Bryant Molecular and genetic methods will be used to investigate the functions of a newly discovered family of Imaginal Disc Growth Factors (IDGFs) from Drosophila. The factors were discovered in conditioned medium from imaginal disc cultures, and have been shown to be related to chitinases rather than to previously known growth factors. They cooperate with insulin to stimulate the proliferation, polarization and motility of imaginal disc cells in vitro. The first goal is to identify the functions of IDGFs in vivo by isolating and characterizing mutations and deficiencies in the genes encoding these proteins. P-element mutagenesis and radiation mutagenesis will be used to produce loss-of-function mutations and deficiencies of the IDGF genes, and the mutant phenotypes produced by single and multiple losses will be analyzed. Genetic methods will also be used to determine whether individual IDGFs can compensate for loss of other members of the family, and to test the hypothesis that these proteins are secreted by the fat body into the blood. The second goal is to test the effects of each IDGF and of combinations of IDGFs, with and without insulin, on cell proliferation and polarization both in cell lines and in larval tissues. Each IDGF will be produced in a baculoviral expression system. The binding of IDGFs to cells and to proteins will be studied in order to determine whether these proteins interact with a cell-surface receptor, and the possibility that they interact directly with insulin will be tested. The IDGFs will also be tested for chitinase activity. The third goal is to test the hypothesis that the expression of IDGF genes is under the control of the known transcription factor serpent, which has an appropriate expression pattern and DNA-binding properties. The discovery of these proteins opens up a variety of new approaches to the problem of growth control in Drosophila.

Peter J. Bryant 分子和遗传学方法将用于研究果蝇中新发现的成虫盘生长因子 (IDGF) 家族的功能。这些因子是在成虫盘培养物的条件培养基中发现的，并已被证明与几丁质酶有关，而不是与先前已知的生长因子有关。它们与胰岛素配合，在体外刺激成虫盘细胞的增殖、极化和运动。第一个目标是通过分离和表征编码这些蛋白质的基因中的突变和缺陷来鉴定 IDGF 的体内功能。将使用P元件诱变和辐射诱变产生IDGF基因的功能丧失性突变和缺陷，并分析单次和多次丧失产生的突变表型。遗传方法还将用于确定个体IDGF是否可以补偿家族其他成员的损失，并检验这些蛋白质是由脂肪体分泌到血液中的假设。第二个目标是测试每种 IDGF 以及 IDGF 组合（有或没有胰岛素）对细胞系和幼虫组织中细胞增殖和极化的影响。每个 IDGF 将在杆状病毒表达系统中产生。将研究 IDGF 与细胞和蛋白质的结合，以确定这些蛋白质是否与细胞表面受体相互作用，并测试它们直接与胰岛素相互作用的可能性。 IDGF 还将测试几丁质酶活性。第三个目标是检验 IDGF 基因的表达受到已知转录因子蛇的控制的假设，蛇具有适当的表达模式和 DNA 结合特性。这些蛋白质的发现为解决果蝇生长控制问题开辟了多种新方法。