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Volume 13
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10.3390/biom13040586
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Article

Proteasome-Associated Proteins, PA200 and ECPAS, Are Essential for Murine Spermatogenesis

by
Ban Sato
1,2,*,
Jiwoo Kim
3,
Kazunori Morohoshi
2,
Woojin Kang
4,
Kenji Miyado
4,
Fuminori Tsuruta
1,
Natsuko Kawano
2 and
Tomoki Chiba
1,*
1
Master’s and Doctoral Program in Biology, Faculty of Life and Environmental Sciences, University of Tsukuba, 1-1-1 Tennodai, Tsukuba 305-8577, Japan
2
Laboratory of Regulatory Biology, Department of Life Sciences, School of Agriculture, Meiji University, 1-1-1 Higashimita, Kawasaki 214-8571, Japan
3
College of Biological Sciences, School of Life and Environmental Sciences, University of Tsukuba, 1-1-1 Tennodai, Tsukuba 305-8577, Japan
4
Department of Reproductive Biology, National Research Institute for Child Health and Development, 2-10-1 Okura, Setagaya 157-8535, Japan
*
Authors to whom correspondence should be addressed.
Biomolecules 2023, 13(4), 586; https://doi.org/10.3390/biom13040586
Submission received: 22 February 2023 / Revised: 16 March 2023 / Accepted: 22 March 2023 / Published: 24 March 2023
(This article belongs to the Special Issue Gametogenesis and Gamete Interaction)
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Abstract

Proteasomes are highly sophisticated protease complexes that degrade non-lysosomal proteins, and their proper regulation ensures various biological functions such as spermatogenesis. The proteasome-associated proteins, PA200 and ECPAS, are predicted to function during spermatogenesis; however, male mice lacking each of these genes sustain fertility, raising the possibility that these proteins complement each other. To address this issue, we explored these possible roles during spermatogenesis by producing mice lacking these genes (double-knockout mice; dKO mice). Expression patterns and quantities were similar throughout spermatogenesis in the testes. In epididymal sperm, PA200 and ECPAS were expressed but were differentially localized to the midpiece and acrosome, respectively. Proteasome activity was considerably reduced in both the testes and epididymides of dKO male mice, resulting in infertility. Mass spectrometric analysis revealed LPIN1 as a target protein for PA200 and ECPAS, which was confirmed via immunoblotting and immunostaining. Furthermore, ultrastructural and microscopic analyses demonstrated that the dKO sperm displayed disorganization of the mitochondrial sheath. Our results indicate that PA200 and ECPAS work cooperatively during spermatogenesis and are essential for male fertility.
Keywords: proteasome; PA200; ECPAS; LPIN1; spermatogenesis; sperm anomalies proteasome; PA200; ECPAS; LPIN1; spermatogenesis; sperm anomalies

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MDPI and ACS Style

Sato, B.; Kim, J.; Morohoshi, K.; Kang, W.; Miyado, K.; Tsuruta, F.; Kawano, N.; Chiba, T. Proteasome-Associated Proteins, PA200 and ECPAS, Are Essential for Murine Spermatogenesis. Biomolecules 2023, 13, 586. https://doi.org/10.3390/biom13040586

AMA Style

Sato B, Kim J, Morohoshi K, Kang W, Miyado K, Tsuruta F, Kawano N, Chiba T. Proteasome-Associated Proteins, PA200 and ECPAS, Are Essential for Murine Spermatogenesis. Biomolecules. 2023; 13(4):586. https://doi.org/10.3390/biom13040586

Chicago/Turabian Style

Sato, Ban, Jiwoo Kim, Kazunori Morohoshi, Woojin Kang, Kenji Miyado, Fuminori Tsuruta, Natsuko Kawano, and Tomoki Chiba. 2023. "Proteasome-Associated Proteins, PA200 and ECPAS, Are Essential for Murine Spermatogenesis" Biomolecules 13, no. 4: 586. https://doi.org/10.3390/biom13040586

APA Style

Sato, B., Kim, J., Morohoshi, K., Kang, W., Miyado, K., Tsuruta, F., Kawano, N., & Chiba, T. (2023). Proteasome-Associated Proteins, PA200 and ECPAS, Are Essential for Murine Spermatogenesis. Biomolecules, 13(4), 586. https://doi.org/10.3390/biom13040586

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