Elongin B is a binding partner of the male germ cell nuclear speckle protein sperm-associated antigen 16S (SPAG16S) and is regulated post-transcriptionally in the testis
Zhengang Zhang A B , Qian Huang C D , Zhenyu Wang C E , Jie Zou F , Zuoren Yu G , Jerome F. Strauss
+ Author Affiliations
- Author Affiliations
A Department of Gastroenterology, Tongji Hospital, Huazhong University of Science and Technology, 1095 Jiefang da dao, Wuhan, Hubei 430030, China.
B Department of Obstetrics and Gynecology, Virginia Commonwealth University, 1101 E. Marshall Street, Richmond, VA 23298, USA.
C Department of Physiology, Wayne State University, 275 E Hancock Street, Detroit, MI 48201, USA.
D Department of Occupational and Environmental Health, Hubei Province Key Laboratory of Occupational Hazard Identification and Control, School of Public Health, Wuhan University of Science and Technology, 2 Huangjiahu xi lu, Wuhan, Hubei 430060, China.
E Department of Biochemistry, School of Medicine, Wuhan University of Science and Technology, 2 Huangjiahu xi lu, Wuhan, Hubei 430065, China.
F Wuhan Institute of Skin Disease Prevention and Control, 64 Wusheng lu, Wuhan, Hubei 430030, China.
G Research Center for Translational Medicine, Tongji University School of Medicine, Shanghai East Hospital, 150 Jimo lu, Shanghai, China.
H Department of Obstetrics and Gynecology, Wayne State University, 275 E Hancock Street, Detroit, MI 48201, USA.
I Corresponding author. Email: gn6075@wayne.edu
Reproduction, Fertility and Development 31(5) 962-971 https://doi.org/10.1071/RD18303
Submitted: 14 August 2018 Accepted: 21 December 2018 Published: 28 February 2019
Abstract
In this study we identified Elongin B, a regulatory subunit of the trimeric elongation factor Elongin ABC, which increases the overall rate of elongation by RNA polymerase II, as a major binding partner of sperm-associated antigen 16S (SPAG16S), a component of nuclear speckles. Nuclear speckles are nuclear subcompartments involved in RNA maturation. Previously, we showed that SPAG16S is essential for spermatogenesis. In the present study, a specific antibody against mouse Elongin B was generated and reacted with a protein with the predicted size of Elongin B in the testis; immunofluorescence staining revealed that the Elongin B was located in the nuclei and residual bodies. In round spermatids, Elongin B was colocalised with splicing factor SC35 (SC35), a marker of nuclear speckles. During the first wave of spermatogenesis, Elongin B transcripts were initially detected at Postnatal Day (PND) 8, and levels were greatly increased afterwards. However, Elongin B protein was only found from PND30, when germ cells progressed through spermiogenesis. Polysomal gradient analysis of Elongin B transcripts isolated from adult mouse testes revealed that most of the Elongin B mRNA was associated with translationally inactive, non-polysomal ribonucleoproteins. An RNA electrophoretic mobility shift assay demonstrated that the 3′ untranslated region of the Elongin B transcript was bound by proteins present in testis but not liver extracts. These findings suggest that post-transcriptional regulation of Elongin B occurs in the testis, which is a common phenomenon during male germ cell development. As a major binding partner of SPAG16S, Elongin B may play an important role in spermatogenesis by modulating RNA maturation.
Additional keywords: 3′ untranslated region, post-transcriptional regulation, spermiogenesis, transcription elongation factor B.
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