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Actions of Seminal Proteins in Mated Drosophila Females

精液蛋白在交配果蝇雌性中的作用

基本信息

批准号：
7346974
负责人：
Mariana Federica Wolfner
金额：
$ 34.27万
依托单位：
CORNELL UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2000
资助国家：
美国
起止时间：
2000-08-01 至 2009-12-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7346974
关键词：
Animals Area Behavior Binding Binding Proteins Biochemical Biological Assay Biological Models Biological Process Blood Circulation Class Cloning Collection Conflict (Psychology)Dissection Drosophila genus Drosophila inturned protein Ectopic Expression Employee Strikes Endopeptidases Evolution Expressed Sequence Tags Female Fertility Fertilization Funding Gene Proteins Gene Targeting Genes Genetic Genome Genomics Gland Goals Hormones Human Individual Insect Vectors Insecta Learning Longevity Mammals Mediating Modeling Modification Molecular Molecular Genetics Names Organism Ovulation Partner in relationship Peptide Hydrolases Peptides Play Process Production Prostate Protease Inhibitor Proteins Proteolysis Range Regulation of Proteolysis Reproduction Reproductive Behavior Research Research Personnel Role Running Screening procedure Seminal Seminal fluid Signal Pathway Signal Transduction Site Sterility Structure System Taxon Techniques Testing Tissues Toxic effect Transgenic Organisms Trypsin Inhibitors base egg fly functional genomics human disease in vitro Assay in vivo insight interest male member practical application programs receptor reproductive reproductive function reproductive hormone research study response sperm cell spermadhesin trait

项目摘要

In animals with internal fertilization, seminal proteins transferred with sperm play crucial roles in the fertility of both males and females. This is best understood in the model system Drosophila, whose seminal proteins ("Acps") induce females to increase egg production, store sperm, and modify their behavior, but also to die younger than unmated females. We have identified 90% of all Acps in the fly genome (52 genes). They include members of conserved seminal-protein classes such as peptides, sperm-binding proteins and protease inhibitors; 29% show signs of having evolved rapidly, a trait seen for reproductive proteins in many organisms. We have also identified effectors and genes regulated by Acps in mated females. Our goal is to determine the Acps that mediate specific reproductive changes in females, and how they carry out their functions. In Aim 1 we will screen for the reproductive function of individual Acps. We will assess the targeting, biochemical activity and structure of these molecules. We will determine their in vivo functions by ectopic expression assays in females, and by the consequences of removing individual Acps from males. We have adapted the Gateway cloning system to carry out these assays with high efficiency. In Aim 2 we will analyze in detail the functions of individual Acps that regulate egg production, sperm storage or lifespan effects/protease inhibition. Initially we will extend our analysis of three Acps, each representing one of these areas. We will determine molecular requirements for their activity, the downstream genes/effectors they regulate, and their binding proteins. We will then similarly analyze Acps identified in Aim 1 to be important in egg production, sperm storage or proteolysis regulation. Through this revised research plan we will obtain a comprehensive functional view of the fly tissue analogous to the mammalian prostate gland, and a specific view of the actions of individual seminal proteins and the responses they trigger in females. This information will help us understand the molecular interactions between males and females, and to evaluate hypotheses about the functions and evolution of reproductive proteins. In addition to their relevance to reproductive mechanisms in a range of organisms including mammals, we anticipate that our findings will have practical applications in the control of insect vectors of human disease.

在体内受精的动物中，随精子转移的精液蛋白在雄性和雌性的生育能力中都起着至关重要的作用。这在模型系统果蝇中得到了最好的理解，果蝇的精液蛋白（“Acps”）诱导雌性增加卵子产量，储存精子，改变它们的行为，但也比未交配的雌性死得更早。我们已经鉴定了果蝇基因组中90%的Acps（52个基因）。它们包括保守的精蛋白类的成员，如多肽、精子结合蛋白和蛋白酶抑制剂；29%显示出快速进化的迹象，这是许多生物体中生殖蛋白的特征。我们还发现了Acps在交配雌性中调控的效应物和基因。我们的目标是确定介导雌性特定生殖变化的Acps，以及它们如何发挥作用。