title: Identification of Ube2b as a novel target of androgen receptor in mouse sertoli cells.
abstract: Many genes are regulated by androgen and its receptor (AR), but the direct target genes of AR, especially those involved in spermatogenesis and male infertility, remain unclear. Here, we identified ubiquitin-conjugating enzyme E2B (Ube2b) as a critical target gene of AR. The expression of UBE2B was decreased in the testes of Sertoli cell AR knockout (S-AR(-/y)) mice analyzed by quantitative RT-PCR (qRT-PCR) and immunofluorescence. The upregulation of Ube2b gene by testosterone was further demonstrated by Western blot and qRT-PCR in TM4 cells, a mouse Sertoli cell line. Moreover, luciferase assay, electrophoretic mobility shift assay, and chromatin immunoprecipitation assay validated that the ligand-bound AR activated Ube2b transcription via direct binding to the androgen-responsive element of the Ube2b promoter. In vitro analyses showed that testosterone increased UBE2B expression and activated H2A ubiquitylation, while downregulation of UBE2B blocked the testosterone-induced H2A ubiquitylation. The ubiquitylation of H2A was markedly decreased in the testes of S-AR(-/y) mice by immunohistochemistry. Digital gene expression analysis showed that 113 genes were significantly downregulated and 71 were upregulated by UBE2B in TM4 cells. These results suggest that Ube2b, as a direct AR transcriptional target in Sertoli cells, mediates the function of AR in spermatogenesis by promoting H2A ubiquitylation.
authors.affiliation: Guangdong and Shenzhen Key Laboratory of Male Reproductive Medicine and Genetics, Institute of Urology, Peking University Shenzhen Hospital, Shenzhen Peking University-Hong Kong University of Science & Technology Medical Center, Shenzhen, China.
authors.forename: Lisha
authors.initials: L
authors.lastname: Mou
authors.forename: Qiaoxia
authors.initials: Q
authors.lastname: Zhang
authors.forename: Yadong
authors.initials: Y
authors.lastname: Wang
authors.forename: Qiang
authors.initials: Q
authors.lastname: Zhang
authors.forename: Liang
authors.initials: L
authors.lastname: Sun
authors.forename: Cailing
authors.initials: C
authors.lastname: Li
authors.forename: Weiren
authors.initials: W
authors.lastname: Huang
authors.forename: Yongxian
authors.initials: Y
authors.lastname: Yuan
authors.forename: Yonggang
authors.initials: Y
authors.lastname: Duan
authors.forename: Ruiying
authors.initials: R
authors.lastname: Diao
authors.forename: Zhimao
authors.initials: Z
authors.lastname: Jiang
authors.forename: Jiongxian
authors.initials: J
authors.lastname: Ye
authors.forename: Zhiming
authors.initials: Z
authors.lastname: Cai
authors.forename: Yaoting
authors.initials: Y
authors.lastname: Gui
mesh_terms: D000818:Animals; D002478:Cells, Cultured; D015536:Down-Regulation / Q000502:physiology; D005786:Gene Expression Regulation; D008297:Male; D051379:Mice; D018345:Mice, Knockout; D011944:Receptors, Androgen / Q000235:genetics / Q000378:metabolism*; D012708:Sertoli Cells / Q000378:metabolism*; D015398:Signal Transduction / Q000502:physiology; D013091:Spermatogenesis / Q000502:physiology*; D044763:Ubiquitin-Conjugating Enzymes / Q000235:genetics / Q000378:metabolism*; D054875:Ubiquitination / Q000502:physiology; D015854:Up-Regulation / Q000502:physiology
chemical_list: D011944:Receptors, Androgen; C465394:Ube2b protein, mouse; D044763:Ubiquitin-Conjugating Enzymes
keywords: Sertoli cells; UBE2B; androgen; androgen receptor; male infertility; spermatogenesis