Actions of Seminal Proteins in Mated Drosophila Females

在体内受精的动物中，与精子一起转移的精液蛋白在 男性和女性的生育能力。这一点在果蝇的模型系统中得到了最好的理解，它的种子 蛋白质(“ACP”)诱导雌性增加产卵量，储存精子，并改变她们的行为，但也 比未交配的雌性更早死亡。我们已经确定了苍蝇基因组(52个基因)中90%的ACPs。他们 包括保守精液蛋白类成员，如多肽、精子结合蛋白和蛋白酶 抑制物；29%的人表现出快速进化的迹象，这是许多生殖蛋白的特征 有机体。我们还在交配的雌性中发现了由ACP调控的效应器和基因。我们的目标是 确定调节雌性特定生殖变化的ACP，以及它们如何执行其 功能。 在Aim 1WE中，我们将筛选单个ACP的生殖功能。我们将评估目标， 这些分子的生化活性和结构。我们将通过异位来确定它们在体内的功能 在雌性中的表达分析，以及从雄性中移除单个ACP的后果。我们有 采用Gateway克隆系统以高效地进行这些分析。 在目标2中，我们将详细分析单个ACP调节产蛋量的功能， 精子储存或寿命效应/蛋白酶抑制。首先，我们将扩展我们对三个ACP的分析，每个ACP 代表了其中的一个区域。我们将确定它们活性的分子需求，下游 它们调控的基因/效应器，以及它们的结合蛋白。然后，我们将以类似的方式分析 目的1在卵子生产、精子保存或蛋白分解调节中起重要作用。 通过这次修订的研究计划，我们将获得苍蝇组织的全面功能图景 类似于哺乳动物的前列腺癌，并对单个精液蛋白的行为有特殊的看法 以及它们在雌性体内引发的反应。这些信息将有助于我们理解分子间的相互作用 并评估有关生殖功能和进化的假说。 蛋白质。除了它们与一系列生物体的生殖机制有关外，包括 我们预计，我们的发现将在控制昆虫媒介方面有实际应用。 人类疾病。

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 1we 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.