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. ? ? ?
