生殖调控基因C3ORF20的功能调节因子鉴定OA

Objective To identify the regulatory factors associated with the reproduction-related open reading frame gene C3ORF20 and explore its regulatory function in reproduction.Methods The spatiotemporal expression of C3ORF20 was an-alyzed using RNA-seq data from human tissue samples and scRNA-seq data from human testis samples.Bioinformatics was used to identify interacting proteins of C3ORF20 and a protein-protein interaction(PPI)network was constructed.KEGG/GO analysis was conducted to explore the signaling pathways and molecular functions involving these interacting proteins.The co-expression of BIRC6 and C3ORF20 was verified through scRNA-seq analysis and RT-PCR.Co-immunoprecipitati-on(Co-IP)and GST pull-down assays were employed to detect protein interactions and fine interaction domains.Ubiquiti-nation and immunoprecipitation were carried out to analyze the effect of BIRC6 on the stability of C3ORF20.Results C3ORF20 mRNA was specifically expressed in early and mid-round spermatids.Interaction between C3ORF20 and BIRC6 was observed,and BIRC6 interacted with the family with sequence similarity 194(FAM194)domain of C3ORF20 through its ubiquitin-conjugating enzymes(UBCs).BIRC6 ubiquitinated C3ORF20,which results in its instability.Conclusion BIRC6,as a regulator of C3ORF20,regulates its stability through the ubiquitination pathway,thereby affecting its regulato-ry function in reproduction.