Evolutionary studies of mouse seminal fluid protein genes

描述(由申请人提供)：这一高级博士后奖学金申请的目标是在三个领域教育申请人：1)基因组学和2)蛋白质组学，包括此类研究所需的实验室技术和计算机工具，以及3)比较跨杂交区域不同基因导入的选择的相对强度所需的统计方法。这项建议的具体目的是：1.利用蛋白质组学和基因组学方法鉴定小鼠精液中的蛋白质成分。基因组学方法将使用小鼠前列腺EST的现有数据来识别候选基因，蛋白质组学方法将使用高通量LC-MS/MS来直接识别小鼠精液的成分；2.对上述小鼠精液蛋白的候选基因进行测序，用于通过中性理论的统计测试进行分析，以寻找这些基因上选择(阳性和阴性)的特定位点证据，以及b)在野生来源的家蝇和肌肉分枝杆菌种群中寻找这些基因中固定的或几乎固定的多态，用于下文目标4中概述的杂交区研究；3.通过询问进化相似的基因是否落入小鼠、人类和其他哺乳动物基因组的同线区域来鉴定精液蛋白的真正同源基因；4.通过绘制欧洲小鼠肌肉杂交区的导入图来测试特定精液蛋白基因的选择：a)发现在两个亚种之间具有固定或几乎固定的多态的候选基因(即，被目标2a显示为在选择中)(上面的目标2b)；以及b)比较精液蛋白基因跨杂交区导入的倾斜区与相对选择较低的基因(例如，许多核等位酶)和高选择的基因(例如，X和Y染色体多态)的倾向性。 相关性：精液蛋白表现出动态的进化史、显著的正向选择和不同谱系之间的可变选择压力。随着我们对精液成分作用的理解不断加深，我们对影响人类和其他动物生殖成功的因素的理解也会不断加深。这一建议将提供来自啮齿动物和灵长类动物的哺乳动物精液蛋白的比较基因组学观点，这将有助于更好地理解和处理人类涉及不孕不育的问题。此外，精液蛋白的主要来源是前列腺癌，这是男性癌症的常见部位。疾病研究可能受益于选择研究，因为正选择通常与人类疾病基因有关。

DESCRIPTION (provided by applicant): The goal of this senior postdoctoral fellowship application is to educate the applicant in three areas: 1) genomics and 2) proteomics, including both the laboratory techniques and the in silico tools necessary for such studies, and 3) the statistical methodology necessary for comparing the relative strength of selection on different genes introgressing across a hybrid zone. The specific aims of this proposal are: 1. To identify the protein components of mouse seminal fluid using proteomic and genomic approaches. The genomic approach will use existing data on ESTs from mouse prostate to identify candidate genes and the proteomic approach will use high-throughput LC-MS/MS to directly identify components of mouse seminal fluid; 2. To sequence candidate genes for the mouse seminal fluid proteins identified above a) in five different rodent species for analysis by statistical tests of the Neutral Theory to look for site-specific evidence of selection (positive and negative) on these genes and b) in wild-derived M. m. domesticus and M. m. musculus populations to look for fixed or nearly fixed polymorphisms in these genes for use in hybrid zone studies outlined in Aim 4 below; 3. To identify true orthologous genes for seminal fluid proteins by asking whether evolutionary similar genes fall within syntenic regions of mouse, human, and other mammalian genomes; 4. To test for selection on specific seminal fluid protein genes by plotting their introgression across the Mus musculus hybrid zone in Europe: a) to discover candidate genes (i.e., shown to be under selection by Aim 2a) that have a fixed or nearly fixed polymorphism between the two subspecies (Aim 2b above); and b) to compare clines of introgression of seminal fluid protein genes across the hybrid zone for seminal fluid protein genes with clines for genes under relatively low selection (e.g., many nuclear allozymes) and genes under high selection (e.g., X and Y chromosome polymorphisms). Relevance: Seminal fluid proteins show dynamic evolutionary histories, significant positive selection and variable selective pressure between lineages. As our understanding of the roles of seminal fluid components grows, so will our understanding of factors that influence reproductive success in humans and other animals. This proposal will provide a comparative genomics view of mammalian seminal fluid proteins from rodents and primates, which should lead to a better understanding of, and ability to cope with human problems involving infertility. Furthermore, the major source of seminal fluid proteins is the prostate, a common site of male cancer. Disease research may benefit from studies of selection, since positive selection is often associated with human disease genes.