Loss of PMFBP1 Disturbs Mouse Spermatogenesis by Downregulating HDAC3 Expression

• Published in: Journal of assisted reproduction and genetics Vol. 40; no. 8; pp. 1865 - 1879
• Main Authors: Xu, Weilong, Yao, Zhoujuan, Li, Yunzhi, Wang, Ke, Kong, Shuai, Wang, Yu, Xiang, Mingfei, Zhu, Fuxi, Wang, Fengsong, Zhang, Hui
• Format: Journal Article
• Language: English
• Published: New York Springer US 01-08-2023; Springer Nature B.V
• Subjects: Animals; Cytoskeletal Proteins - genetics; Flagella; Gamete Biology; Gynecology; Histone deacetylase; Histones; Human Genetics; Immunoblotting; Immunoprecipitation; Male; Mass spectroscopy; Medicine; Medicine & Public Health; Mice; Molecular modelling; Phenotypes; Protein interaction; Proteins; Proteins - metabolism; Proteomics; Reproductive Medicine; RING finger proteins; Semen; Sperm; Spermatogenesis; Spermatogenesis - genetics; Spermatozoa - metabolism; Testis - metabolism; Transcriptomes; Ubiquitin-Protein Ligases; CCT3; Proteome; HDAC3; PMFBP1; Protein-protein interaction; Spermatogenesis
• ISSN: 1058-0468; 1573-7330; 1573-7330
• DOI: 10.1007/s10815-023-02874-0

Purpose
Polyamine modulating factor 1 binding protein (PMFBP1) acts as a scaffold protein for the maintenance of sperm structure. The aim of this study was further to identify the new role and molecular mechanism of PMFBP1 during mouse spermatogenesis.
Methods and Results
We identified a profile of proteins interacting with PMFBP1 by immunoprecipitation combined with mass spectrometry and demonstrated that class I histone deacetylases, particularly HDAC3 and chaperonin-containing TCP1 subunit 3 (CCT3), were potential interaction partners of PMFBP1 based on network analysis of protein-protein interactions and co-immunoprecipitation. Immunoblotting and immunochemistry assays showed that loss of
Pmfbp1
would result in a decline in HDACs and change the proteomic profile of mouse testis, in which differently expressed proteins are associated with spermatogenesis and assembly of flagella, which was proved by proteomic analysis of testis tissue obtained from
Pmfbp1
−/−
mice. After integrating with transcriptome data for
Hdac3
−/−
and
Sox30
−/−
round sperm obtained from a public database, RT-qPCR confirmed ring finger protein 151 (Rnf151) and ring finger protein 133 (Rnf133) were key downstream response factors of the Pmfbp1-Hdac axis affecting mouse spermatogenesis.
Conclusion
Taken together, this study indicates a previously unidentified molecular mechanism of PMFBP1 in spermatogenesis whereby PMFBP1 interacts with CCT3, affecting the expression of HDAC3, followed by the downregulation of RNF151 and RNF133, resulting in an abnormal phenotype of sperm beyond the headless sperm tails. These findings not only advance our understanding of the function of Pmfbp1 in mouse spermatogenesis but also provide a typical case for multi-omics analysis used in the functional annotation of specific genes.