Evolutionary rate covariation identifies new members of a protein network required for Drosophila melanogaster female post-mating responses.
Evolutionary rate covariation identifies new members of a protein network required for Drosophila melanogaster female post-mating responses.
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DOI：
10.1371/journal.pgen.1004108
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发表时间：
2014-01
期刊：
影响因子：
4.5
通讯作者：
Wolfner MF
中科院分区：
文献类型：
--
作者：
Findlay GD;Sitnik JL;Wang W;Aquadro CF;Clark NL;Wolfner MF
在交配过程中从雄性转移到雌性的精液蛋白质是完全生育所必需的，并且可以对雌性的生理和行为产生显着的影响。在黑腹果蝇中，精液蛋白性肽（SP）通过增加产卵量和降低对求爱的接受性来影响交配的雌性。这些行为变化会持续几天，因为SP与储存在雌性体内的精子结合。然后SP逐渐释放，使其与其雌性表达的受体相互作用。SP与精子的结合需要五种额外的精液蛋白，它们在网络中共同作用。在这个物种中已经鉴定出数百种未表征的雄性和雌性蛋白质，但是单独筛选每种蛋白质的网络功能将带来后勤挑战。为了优先筛选这些蛋白质参与SP网络，我们使用了比较基因组学方法，以确定候选蛋白质的进化速率在整个果蝇胚胎发育协变与SP网络蛋白。随后的功能测试的18个共同变化的候选人通过RNA干扰确定了三个男性精液蛋白和三个女性生殖道蛋白，每个所需的长期持久性的SP反应的女性。分子遗传分析表明，这三种新的雄性蛋白质是将其他网络蛋白质转移到雌性以及SP与储存在交配雌性中的精子结合所必需的。相反，这三种雌性蛋白在SP结合和精子储存的下游起作用。这些发现扩大了SP在女性中的作用所需的精液蛋白质的数量，并表明多种女性蛋白质是SP反应所必需的。此外，我们的功能分析表明，进化速率协变是一个有价值的预测工具，用于识别相互作用的蛋白质网络的候选成员。生殖需要的不仅仅是精子和卵子。在体内受精的动物中，精液和雌性中的其他蛋白质对完全生育力至关重要。尽管已知有数百种这样的生殖蛋白，但我们了解它们如何相互作用的能力仍然有限。在这项研究中，我们研究了是否共享的蛋白质序列进化模式的预测功能的相互作用，集中在一个小的网络的蛋白质，控制生育能力和女性交配后的行为，果蝇。我们首先发现，已知在这个网络中起作用的六种蛋白质显示了果蝇遗传学中相关的进化模式。然后，我们筛选了数百种其他未知的雄性和雌性生殖蛋白，并确定了那些与已知网络蛋白进化模式最相似的蛋白。我们测试了这些候选基因中的每一个，并发现了六个新的网络成员，每个成员都是长期生育所必需的。利用分子遗传学，我们还观察到，这些新蛋白质作用的网络步骤与它们最强的进化相关性一致。我们的研究结果表明，模式的共同进化可能是广泛有用的预测蛋白质相互作用的各种生物过程。
Seminal fluid proteins transferred from males to females during copulation are required for full fertility and can exert dramatic effects on female physiology and behavior. In Drosophila melanogaster, the seminal protein sex peptide (SP) affects mated females by increasing egg production and decreasing receptivity to courtship. These behavioral changes persist for several days because SP binds to sperm that are stored in the female. SP is then gradually released, allowing it to interact with its female-expressed receptor. The binding of SP to sperm requires five additional seminal proteins, which act together in a network. Hundreds of uncharacterized male and female proteins have been identified in this species, but individually screening each protein for network function would present a logistical challenge. To prioritize the screening of these proteins for involvement in the SP network, we used a comparative genomic method to identify candidate proteins whose evolutionary rates across the Drosophila phylogeny co-vary with those of the SP network proteins. Subsequent functional testing of 18 co-varying candidates by RNA interference identified three male seminal proteins and three female reproductive tract proteins that are each required for the long-term persistence of SP responses in females. Molecular genetic analysis showed the three new male proteins are required for the transfer of other network proteins to females and for SP to become bound to sperm that are stored in mated females. The three female proteins, in contrast, act downstream of SP binding and sperm storage. These findings expand the number of seminal proteins required for SP's actions in the female and show that multiple female proteins are necessary for the SP response. Furthermore, our functional analyses demonstrate that evolutionary rate covariation is a valuable predictive tool for identifying candidate members of interacting protein networks. Reproduction requires more than a sperm and an egg. In animals with internal fertilization, other proteins in the seminal fluid and the female are essential for full fertility. Although hundreds of such reproductive proteins are known, our ability to understand how they interact remains limited. In this study, we investigated whether shared patterns of protein sequence evolution were predictive of functional interactions by focusing on a small network of proteins that control fertility and female post-mating behavior in the fruit fly, Drosophila melanogaster. We first showed that the six proteins already known to act in this network display correlated patterns of evolution across the Drosophila phylogeny. We then screened hundreds of otherwise uncharacterized male and female reproductive proteins and identified those with patterns of evolution most similar to those of the known network proteins. We tested each of these candidate genes and found six new network members that are each required for long-term fertility. Using molecular genetics, we also observed that the steps in the network at which these new proteins act are consistent with their strongest evolutionary correlations. Our results suggest that patterns of coevolution may be broadly useful for predicting protein interactions in a variety of biological processes.
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