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Location: Cool and Cold Water Aquaculture Research

Title: Fbos, a novel oocyte-specific protein, interacts with proteins important for oocyte development in rainbow trout (Oncorhynchus mykiss)

item Wang, Lei
item Tripurani, Swamy
item Wanna, Warapond
item Rexroad, Caird
item Yao, Jianbo

Submitted to: Society for the Study of Reproduction Annual Meeting
Publication Type: Abstract Only
Publication Acceptance Date: 4/9/2010
Publication Date: 7/30/2010
Citation: Wang, L., Tripurani, S.K., Wanna, W., Rexroad III, C.E., Yao, J. 2010. Fbos, a novel oocyte-specific protein, interacts with proteins important for oocyte development in rainbow trout (Oncorhynchus mykiss) [abstract]. Society for the Study of Reproduction Annual Meeting. Paper No. 410.

Interpretive Summary:

Technical Abstract: Oogenesis is characterized by a series of developmentally regulated events, which result in the matured oocyte that can give rise to a new organism after fertilization. Oocyte-specific genes play critical roles in oogenesis; however, the molecular details of oocyte-specific genes are poorly defined. Through analysis of expressed sequence tags (ESTs) from a rainbow trout oocyte cDNA library, we identified and characterized a novel oocyte-specific gene coding for an F-box protein (Fbos, F-box oocyte-specific). F-box proteins are components of ubiquitin-ligase complexes, which bind substrates for uiquitin-mediated proteolysis. F-box proteins have also been associated with cellular functions such as signal transduction and regulation of the cell cycle. To define the molecular mechanisms of Fbos-dependent biological processes during oogenesis in rainbow trout, we used Fbos as bait in a yeast two-hybrid screen system, along with a rainbow trout ovary cDNA library. More than 300 yeast colonies were able to grow on nutritional selective plates; 18 clones displayed strong LacZ activity as shown by colony lift filter assay for '-galactosidase activity. After verifying the interactions by reintroduction of the candidate prey plasmids into yeast reporter strain containing Fbos plasmid, 8 plasmids were rescued successfully and subjected to DNA sequence analysis. The sequencing data demonstrated that Fbos protein interacts with a number of proteins including histone H3.3, adenylate kinase 2, ribosomal protein L36, oocyte protease inhibitor 1, zona pellucida protein 2 (ZP2) and tissue inhibitor of metalloproteinase 2 (TIMP-2). All 5 proteins showed significant '-galactosidase activities at different levels compared to the control in a quantitative ß-galactosidase assay, indicating different binding affinities of these proteins with Fbos protein. Some of these proteins like ZP2 and TIMP-2 are known to play critical roles in oocyte development in various species. Our results suggest that through protein-protein interactions, the novel oocyte-specific F-box protein may play an important role in early oocyte development by regulating other critical proteins involved in oogenesis in rainbow trout.