Maternal control of the Drosophila embryonic Dorsal-Ventral axis

果蝇胚胎背腹轴的母体控制

• 批准号: 7316772
• 负责人: DAVID S. STEIN
• 金额: $ 27.46万
• 依托单位: UNIVERSITY OF TEXAS AT AUSTIN
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-08-15 至 2011-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7316772
• 关键词: Amino Acid Sequence; Area; Blood Clot; Blood coagulation; Cause of Death; Cells; Cleaved cell; Data; Dependence; Development; Disease; Dorsal; Dorsal-Ventral Pattern Formation; Drosophila genus; Embryo; Endopeptidases; Environment; Etiology; Event; Exhibits; Female; Fibrinolysis; Gene Expression; Generations; Genes; Genetic; Genetic Screening; Glycolipids; Glycoproteins; Glycosaminoglycans; Goals; Health; Human; Human Resources; Inorganic Sulfates; Institutes; Invaded; Knowledge; Laboratories; Lead; Ligands; Localized; Membrane; Microscopy; Mission; Modeling; Molecular; Molecular and Cellular Biology; Mutation; Myocardial Infarction; Oogenesis; Pathway interactions; Pattern; Peptide Hydrolases; Perivitelline Space; Physiological Processes; Process; Production; Protein Isoforms; Proteins; Regulation; Research; Research Personnel; Resources; Role; Serine; Serine Protease; Side; Site; Spatial Distribution; Specificity; Stroke; Students; Testing; Texas; Therapeutic Agents; United States National Institutes of Health; Universities; Unspecified or Sulfate Ion Sulfates; Work; base; burden of illness; complement pathway; design; egg; experience; improved; insight; microorganism; mutant; protein protein interaction; receptor; research study; sulfation; sulfotransferase

DESCRIPTION (provided by applicant): Dorsal-ventral polarity in the Drosophila embryo is initiated during oogenesis by expression of the Pipe sulfotransferase in the ventral follicle cells which, by an unknown mechanism, leads to the localized activation of a serine protease cascade in the perivitelline space between the embryonic membrane and the eggshell. Our long-term goal is to understand how this serine protease cascade is spatially regulated. The objectives of this application are to elucidate the mechanism through which spatial restriction of the serine protease cascade is implemented and to identify the target of Pipe action in the follicle cell layer. Our central hypothesis is that Pipe activity in the ventral follicle cells results in the sulfation of a glycoprotein or glycolipid that is secreted into and localized within the ventral perivitelline space where it brings about the localized activation of the serine protease cascade. The rationale for the proposed research is that it will greatly expand our understanding of DV pattern formation and will also provide insight into mechanisms that regulate analogous localized serine proteolytic events that influence human health such as the ones involved in the formation and breakdown of blood clots. Blood coagulation and fibrinolysis are critical factors in the etiology of thrombotic diseases such as heart attack and stroke which represent leading causes of death worldwide. Thus, the proposed research is relevant to the mission of the NIH to obtain fundamental knowledge that will potentially reduce the burden of disease. We will pursue two Specific Aims: 1) To determine the extent to which the activity of the serine protease cascade is regulated by the localization of the proteases themselves, by their spatially restricted activation or activities, or by physical interactions with one another that modulate their activities. Functional, tagged versions of the proteases will be used to examine their processing and spatial distribution and to investigate protein-protein interactions in wildtype and various genetic backgrounds. 2) To identify genes involved in the synthesis of the target of Pipe sulfotransferase activity in the ventral follicle cells. Genetic screens will be carried out to identify mutations that lead to the production of dorsalized embryos by mutant females. The proposed research is significant as it will yield new insights into mechanisms that regulate serine protease action and thus may contribute to the development of improved therapeutic agents for the modulation of serine proteases that influence human health.

描述(由申请人提供)：果蝇胚胎的背腹两极是在卵子发生过程中通过腹侧毛囊细胞中管道磺基转移酶的表达而启动的，这种表达以一种未知的机制导致在胚膜和蛋壳之间的卵周隙中局部激活丝氨酸蛋白酶级联。我们的长期目标是了解这种丝氨酸蛋白酶级联是如何在空间上受到调控的。这项应用的目的是阐明丝氨酸蛋白酶级联的空间限制的实现机制，并确定毛囊细胞层中管道作用的靶点。我们的中心假设是腹侧毛囊细胞中的PIPE活性导致糖蛋白或糖脂的硫酸盐化，该糖蛋白或糖脂被分泌并定位于腹侧卵周间隙，在那里它导致丝氨酸蛋白酶级联的局部激活。这项研究的基本原理是，它将极大地扩展我们对DV模式形成的理解，并将为调节影响人类健康的类似局部丝氨酸蛋白分解事件的机制提供洞察力，例如参与血栓形成和破裂的那些事件。凝血和纤溶是心脏病和中风等血栓性疾病病因学的关键因素，这些疾病是全世界主要的死亡原因。因此，拟议的研究与美国国立卫生研究院获取基础知识的使命相关，这些基础知识可能会减少疾病的负担。我们将追求两个特定的目标：1)确定丝氨酸蛋白酶级联的活性在多大程度上受到蛋白酶本身的定位、它们空间上受限的激活或活性、或相互之间调节其活性的物理相互作用的调节。这些酶的功能性标记版本将被用来检查它们的加工和空间分布，并研究野生型和不同遗传背景下的蛋白质-蛋白质相互作用。2)寻找腹侧毛囊细胞合成导管磺基转移酶靶标的相关基因。将进行基因筛查，以确定导致突变雌性产生背部胚胎的突变。这项拟议的研究具有重要意义，因为它将对调节丝氨酸蛋白酶作用的机制产生新的见解，从而可能有助于开发更好的治疗药物来调节影响人类健康的丝氨酸蛋白酶。