SUMO1 modification reduces oxidative stress and SUMO1ylated AKAP4 degradation affects frozen-thawed boar sperm quality.

Abstract

Low-temperature injury affects normal physiological function and viability of boar sperm during cryopreservation. Small ubiquitin-like modifier (SUMO) modification of proteins after translation is related to the cell stress response but the relationship between SUMO modification and oxidative stress in freeze-thawed sperm remains unclear. A-kinase ankyrin 4 (AKAP4) and its precursor proAKAP4 are two main proteins in mammalian sperm. Although AKAP4 expression has been studied in many species, its expression in porcine sperm has not been described in detail. In this study, liquid chromatography-mass spectrometry was used to determine the differentially expressed SUMO-modified proteins in porcine sperm after freezen and thawed. The results identified 26 down-regulated SUMO-modified proteins, with AKAP4 identified as one of the target proteins of SUMO1 under sperm stress. In addition, the level of SUMO1 protein increased significantly (P < 0.001) and the level of AKAP4 protein decreased (P < 0.05) after freezing and oxidative stress treatment. Inhibition of SUMO1 modification of AKAP4 protein did not affect its degradation (P > 0.05), indicating that SUMO1 is not involved in the degradation of AKAP4. The inhibition of SUMO1 modification by sperm protein decreased sperm motility (P < 0.05), ATP content, and DNA integrity (P < 0.05). In summary, cryopreservation and oxidative stress can induce SUMO modification of porcine sperm proteins and the modification of sperm protein SUMO1 can help sperm resist oxidative stress; and its role in protecting sperm quality is not via regulating the degradation of AKAP4. These findings provide new insights into the mechanisms underlying SUMO1 modifications during sperm cryopreservation and oxidative stress.