Objective: S-sulfhydration of proteins is reported to be an important downstream event in signaling mediated by H 2 S, a crucial endogenous gasotransmitter. However, studies on the role of S-sulfhydrated proteins in the ovaries are limited. In this study, we sought to identify and study the function and mechanism of S-sulfhydration of histone variant H3.3 (sH3.3) in mouse granulosa cells. Methods: The levels of sH3.3, indicated enzymes and H 2 S in the primary granulosa cells or KGN cell line The Biotin-HPDP switch, western blotting and Endogenous H 2 S determination were performed to determine. Ovary tissues and serum were collected from the H3f3b C111S mutant and wildtype control mice. RT-PCR, immunofluorescence, IHC assay, western blotting, TUNEL, MDA, and ROS assays were employed to determine the function of sH3.3. The Co-immunoprecipitation (Co-IP), Electrophoretic mobility shift, and Luciferase reporter assays were used to the working mechanism of the sH3.3. Results: Using the mouse model, we showed that sH3.3 deficiency led to female infertility due to ovarian dysfunction. Biological processes, including cell adhesion, steroid biosynthesis, and vascular remodeling, were impaired. Furthermore, we found that sH3.3 deficiency led to reduced ovarian antioxidant capacity. Mechanistically, we demonstrated that sH3.3 deficiency disrupted the interaction with E2F transcription factor 1, which further impaired binding to the Adam19 (a disintegrin and metalloprotease 19) promoter, resulting in reduced Adam19 expression. The subsequent reduction in integrin subunit β1 (ITGB1) shedding led to impairment of the integrin-FAK-ERK1/2 signaling axis, which is critical in granulosa cells. Conclusion: Collectively, our findings reveal a critical role of S-sulfhydration of H3.3 in the ovary for the first time. sH3.3-mediated transcriptional regulation identifies a new mechanism of H 2 S action in mouse granulosa cells.
Chen et al. (Wed,) studied this question.