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BNC1 Promotes Spermatogenesis by Regulating Transcription of Ybx2 and Papolb via Direct Binding to Their Promotor Elements

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Abstract

BNC1 is a transcription factor that is crucial for spermatogenesis and male fertility, although the underlying mechanism remains unclear. To study BNC1’s specific role in spermatogenesis, we characterized a previously developed mouse model carrying a truncating mutation in Bnc1 (termed Bnc1+/tr for heterozygotes and Bnc1tr/tr for homozygotes) and found that the mutation decreased BNC1 protein levels and resulted in germ cell loss by apoptosis. Given that loss of functional Bnc1 is known to result in decreased expression of the spermatogenesis genes Ybx2 and Papolb, we aimed to explore whether and how BNC1 promotes transcription of Ybx2 and Papolb to mediate its role in spermatogenesis. We confirmed significant reduction in YBX2 and PAPOLB protein levels in testis tissue from Bnc1+/tr and Bnc1tr/tr males compared with wild-type mice (Bnc1+/+). Consistently, knockdown of Bnc1 led to downregulation of Ybx2 and Papolb in CRL-2196 cells in vitro. To investigate if BNC1 directly induces Ybx2 and Papolb gene expression, chromatin immunoprecipitation using mouse testicular tissue and luciferase reporter assays in HEK293 cells were used to identify functional binding of BNC1 to the Ybx2 and Papolb promoters at defined BNC1 binding sites. Taken together, this study reveals a mechanism for BNC1’s role in spermatogenesis by directly binding to BNC1 binding elements in the promoter regions of both Ybx2 and Papolb and inducing transcription of these important spermatogenesis genes.

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Abbreviations

BNC1:

Basonuclin 1

YBX2:

Y-box-binding protein 2

PAPOLB:

Poly(A) polymerase beta

BBS:

BNC1 binding sites

SSC:

Spermatogonial stem cell

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Acknowledgments

We thank Professor Sun Fei for supplying cell line C18-4 and Professor Yibing Han for supplying cell line CRL-2196. We thank Professor Sharon YC Ruan from Hongkong Polytechnic University for revising the manuscript.

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Author notes

  1. Jing-Yi Li and Yan-Yun Ying contributed equally to this work.

Authors and Affiliations

  1. Key Laboratory of Reproductive Genetics (Ministry of Education) and Department of Reproductive Endocrinology, Women’s Hospital, Zhejiang University School of Medicine, Hangzhou, 310006, Zhejiang, People’s Republic of China

    Jing-Yi Li, Yan-Yun Ying, Yu-Li Qian, Jian-Peng Chen, Yun Huang, Juan Liu, Ping-Ping Lv, Yi-Feng Liu, Xiao-Ling Hu, Jian-Zhong Sheng, He-Feng Huang & Dan Zhang

  2. Department of Pathology, Harvard Medical School and Brigham and Women’s Hospital, Boston, MA, USA

    Samantha L. P. Schilit

  3. Department of Pathology & Pathophysiology, Zhejiang University School of Medicine, Hangzhou, 310000, Zhejiang, People’s Republic of China

    Jian-Zhong Sheng

  4. International Peace Maternal and Child Health Hospital, Shanghai Jiao Tong University, Shanghai, 200030, People’s Republic of China

    He-Feng Huang

  5. Laboratory for Molecular Medicine, Mass General Brigham Personalized Medicine, Cambridge, MA, USA

    Samantha L. P. Schilit

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  1. Jing-Yi Li
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Li, JY., Ying, YY., Qian, YL. et al. BNC1 Promotes Spermatogenesis by Regulating Transcription of Ybx2 and Papolb via Direct Binding to Their Promotor Elements. Reprod. Sci. 28, 785–793 (2021). https://doi.org/10.1007/s43032-020-00342-z

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