Maternal Control of the Drosophila Embryonic Dorsal/Ventral Axis

果蝇胚胎背/腹轴的母体控制

基本信息

批准号：
8788038
负责人：
DAVID S. STEIN
金额：
$ 30.95万
依托单位：
UNIVERSITY OF TEXAS AT AUSTIN
依托单位国家：
美国
项目类别：
财政年份：
2007
资助国家：
美国
起止时间：
2007-08-15 至 2016-12-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8788038
关键词：
Affect Bacteria Behavior Binding Biological Biological Models Blood Coagulation Disorders Blood coagulation Carbohydrates Cells Cleaved cell Cues Data Development Disease Dorsal Drosophila genus Elements Embryo Epithelium Etiology Future Gene Expression Generations Glycoproteins Glycosaminoglycans Goals Growth Factor Growth Factor Receptors Health Homologous Gene Human Immune Infection Inorganic Sulfates Investigation Knowledge Ligands Lighting Mediating Membrane Mission Molecular Nature Neurons Oocytes Oogenesis Organism Outcome Parasites Parasitic infection Pathway interactions Pattern Peptide Hydrolases Peptides Perivitelline Space Predisposition Process Property Proteins Public Health Receptor Signaling Regulation Research Role Serine Serine Protease Side Signal Pathway Signal Transduction Snakes Structure Testing Therapeutic Unspecified or Sulfate Ion Sulfates Viral Virus Diseases Vitelline Membrane Work base burden of illness cell motility egg gastrulation innovation insight microbial novel receptor sulfotransferase sulfurtransferase vitelline membrane proteins

项目摘要

DESCRIPTION (provided by applicant): Sulfated carbohydrates such as glycosaminoglycans control many, diverse processes affecting human health including blood clotting, immune and neural cell migration, viral and microbial infection, and growth factor/receptor signaling. However, for many of these processes, how the sulfated carbohydrates mediate their biological effects at the mechanistic level is not understood, presenting a barrier to our ability to harness the properties of these molecules for therapeutic purposes. Drosophila embryo dorsal-ventral polarity is determined by a sulfated carbohydrate embedded in the eggshell that influences serine proteolytic activity in the perivitelline space between the embryonic membrane and the eggshell. Gastrulation Defective (GD) processes and activates Snake, Snake processes and activates Easter, and Easter cleaves the Spaetzle precursor into a ligand for the transmembrane receptor Toll. Activation of Easter occurs solely on the ventral side of the egg under the control of Pipe, a homologue of vertebrate glycosaminoglycan sulfating enzymes that is expressed in ventral follicle cells of the developing oocyte. Pipe transfers sulfate to the carbohydrate side chains of glycoproteins that become incorporated into the inner vitelline membrane layer of the eggshell and constitute a ventral cue that promotes processing of Easter by activated Snake. The long-term goal of this project is to determine how the sulfated ventral cue controls the spatially restricted processing of Easter, and in turn, the activation of Toll. Th central hypothesis, which is based on the recent observation that GFP-tagged GD becomes ventrally localized in the perivitelline space, is that processed GD binds to the sulfated ventral cue and once concentrated there, facilitates an interaction between Snake and Easter that results in Easter processing. The proposed investigations pursue three specific aims: 1) To elucidate the mechanism that controls the ventral localization of GD within the perivitelline space of the egg by determining the extent to which GD processing is required for its localization and by identifying determinants within GD that are required for localization and processing. 2) To determine how localized GD mediates the cleavage of Easter by Snake by identifying the GD domains that are required for its interactions with Easter and Snake, and by determining the extent to which GD promotes Snake protease activity, Easter susceptibility to cleavage, or productive interaction between the two proteins. 3) To elucidate the structure of the Pipe-modified carbohydrates that comprise the ventral cue and determine how these molecules facilitate GD localization and Easter processing by isolating VML, a key eggshell protein modified by Pipe, examining its associated carbohydrates by mass spectrometric analysis, and identifying proteins that interact with it in a Pipe/ventral cue-dependent manner. The proposed studies are significant because in the course of elucidating the molecular mechanism underlying this paradigmatic developmental pathway, they will yield new insights into the ways in which serine proteases and other medically important proteins are influenced by their interactions with sulfated carbohydrates.

描述（由申请人提供）：硫化碳水化合物（例如糖胺聚糖）控制着许多影响人类健康的多种过程，包括血液凝结，免疫和神经细胞迁移，病毒和微生物感染以及生长因子/受体信号传导。但是，对于许多此类过程，尚不理解硫酸化的碳水化合物如何在机械水平上介导其生物学作用，这是我们利用这些分子的特性以用于治疗目的的障碍。果蝇胚胎背腹性极性取决于蛋壳中嵌入的硫酸盐碳水化合物，该碳水化合物影响胚胎膜和蛋壳之间的蛋白蛋白蛋白水解活性。胃缺陷（GD）的过程并激活蛇，蛇的过程并激活复活节，复活节将Spaetzle前体切割成跨膜受体Toll的配体。复活节的激活仅发生在管道控制下的卵子的腹侧，这是脊椎动物糖胺聚糖硫化酶的同源物，该酶在发育中的卵母细胞的腹侧卵泡细胞中表达。管道将硫酸盐转移到糖蛋白的碳水化合物侧链中，这些糖蛋白掺入蛋壳的内部卵黄膜层中，并构成一种腹提示，该提示通过活性蛇促进复活节。该项目的长期目标是确定硫酸化的腹cue如何控制复活节的空间限制处理，进而依次激活通行费。基于最近观察到的中央假设，即GFP标记的GD被腹侧定位在Perivitelline空间中，它是处理后的GD与硫酸化的腹cue结合并一旦将其集中在那里，可以促进蛇与复活节之间的相互作用，从而在复活节加工中产生。拟议的调查追求三个具体目的：1）阐明控制GD在卵中的腹膜腹腹腹置定位的机制，通过确定GD加工的定位所需的程度以及确定其定位和加工所需的GD中的确定因素所需的程度。 2）确定如何通过鉴定其与复活节和蛇相互作用所需的GD结构域来介导蛇的裂解，并确定GD在多大程度上促进蛇蛋白酶活性，复活节对两种蛋白质之间的裂解或有效的相互作用的敏感性。 3) To elucidate the structure of the Pipe-modified carbohydrates that comprise the ventral cue and determine how these molecules facilitate GD localization and Easter processing by isolating VML, a key eggshell protein modified by Pipe, examining its associated carbohydrates by mass spectrometric analysis, and identifying proteins that interact with it in a Pipe/ventral cue-dependent manner.拟议的研究很重要，因为在阐明这种范式发育途径的分子机制的过程中，它们将对丝氨酸蛋白酶和其他医学重要的蛋白质的方式产生新的见解。

项目成果

期刊论文数量（8）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Sulfation of eggshell components by Pipe defines dorsal-ventral polarity in the Drosophila embryo.

DOI：
10.1016/j.cub.2009.05.050
发表时间：
2009-07-28
期刊：
Current biology : CB
影响因子：
0
作者：
Zhang Z;Stevens LM;Stein D
通讯作者：
Stein D

No requirement for localized Nudel protein expression in Drosophila embryonic axis determination.

果蝇胚胎轴测定中不需要局部 Nudel 蛋白表达。

DOI：
10.4161/fly.6794
发表时间：
2008
期刊：
Fly
影响因子：
1.2
作者：
Stein,David;Cho,YongSuk;Zhang,Zhenyu;Stevens,LeslieM
通讯作者：
Stevens,LeslieM

A ventrally localized protease in the Drosophila egg controls embryo dorsoventral polarity.

DOI：
10.1016/j.cub.2012.03.065
发表时间：
2012-06-05
期刊：
CURRENT BIOLOGY
影响因子：
9.2
作者：
Cho, Yong Suk;Stevens, Leslie M.;Sieverman, Kathryn J.;Nguyen, Jesse;Stein, David
通讯作者：
Stein, David

Localized serine protease activity and the establishment of Drosophila embryonic dorsoventral polarity.

局部丝氨酸蛋白酶活性和果蝇胚胎背腹极性的建立。