|ZANG, XIAOWEI - West Virginia University|
|MA, HAO - West Virginia University|
|FU, LIYUAN - West Virginia University|
|YAO, JIANBO - West Virginia University|
Submitted to: Society for the Study of Reproduction Annual Meeting
Publication Type: Abstract Only
Publication Acceptance Date: 4/1/2013
Publication Date: 7/22/2013
Citation: Zang, X., Ma, H., Fu, L., Rexroad III, C.E., Yao, J. 2013. Characterization and expression analysis of genes involved in SUMOylation during embryogenesis in rainbow trout (Oncorhynchus mykiss). Society for the Study of Reproduction Annual Meeting. A297.
Technical Abstract: SUMOylation is the post-translational modification of proteins by the addition of the small ubiquitin-like modifier (SUMO), which plays an important role in various cellular processes. It has been reported that SUMO and its related proteins are important in diverse reproductive functions such as ovulation, gametogenesis, and embryogenesis. Modification of target proteins by SUMO is an ATP-dependent enzymatic cascade involving three key enzymes: E1 activating enzyme (the heterodimer SAE1-SAE2), E2 conjugating enzyme (UBC9) and several E3 ligating enzymes (PIAS, RanBP2/Nup358 and Pc2). The objectives of this study were to characterize the genes involved in SUMOylation and determine their expression profiles during embryogenesis in rainbow trout. Through database analysis, we identified 10 Sumo related genes, which include Sumo1, Sumo2, Sumo3, Sae1, Sae2, Ubc9, Pias1, Pias4, Cbx4, and Nse2. Analysis of protein sequences revealed that SUMO1, SUMO2, SUMO3, UBC9, CBX4 and NSE2 are highly conserved at the N terminus, while SAE1 and SAE2 are well conserved at the C terminus among human, mice, zebrafish and rainbow trout species. The expression profiles of the Sumo related genes during embryonic development in rainbow trout were analyzed by quantitative real time PCR using cDNAs derived from unfertilized eggs and embryos of different developmental stages (0h, 3h, 7.5h, 11.5h, 13.5h, 18h, 27h, 34h, 2d, 3d, 4d, 5d, 6d, 8d, 10d, 12d, 16d and 25d post fertilization). The expression of Sumo3, Sae2, Ubc9, Pias4, and Nse2 genes showed similar patterns, being low in unfertilized eggs and increasing gradually in early embryos until 18 h post fertilization followed by a gradual decrease in embryos after 18 h post fertilization; both Sumo1 and Sumo2 genes were highly expressed during and after maternal to zygotic transition; while Sae1, Pias1, and Cbx4 were expressed constitutively at steady-state levels throughout embryogenesis. The data indicate that the expression of Sumo related genes are dynamically regulated during embryonic development in rainbow trout. Current work is focused on analyzing the expression of these genes in eggs of various qualities (post-ovulatory aged eggs) and determining if the oocyte- specific proteins, factor in the germline alpha (Figla) and zygote arrest 1 (Zar1), are sumoylated by in vivo and in vitro assays.