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Published on: September 11, 2014
Loss of PMFBP1 Disturbs Mouse Spermatogenesis by Downregulating HDAC3 Expression
Weilong Xu1, Zhoujuan Yao1, Yunzhi Li1
1School of Life Science, Anhui Medical University, Hefei, 230022, China.
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.
Insights
Polyamine modulating factor 1 binding protein (PMFBP1) interacts with CCT3, impacting HDAC3 expression and leading to abnormal sperm development. This study reveals a new mechanism for PMFBP1 in mouse spermatogenesis.
Area of Science:
- Reproductive Biology
- Molecular Genetics
- Proteomics
Background:
- Polyamine modulating factor 1 binding protein (PMFBP1) is crucial for sperm structure.
- Its precise role and molecular mechanisms in spermatogenesis require further elucidation.
Purpose of the Study:
- To identify novel functions and molecular mechanisms of PMFBP1 in mouse spermatogenesis.
- To investigate the protein interactions and downstream effects of PMFBP1.
Main Methods:
- Immunoprecipitation coupled with mass spectrometry to identify PMFBP1 interacting proteins.
- Co-immunoprecipitation, immunoblotting, and immunochemistry to validate interactions and protein expression.
- Proteomic and transcriptomic analyses of wild-type and Pmfbp1 knockout mouse testes.
- RT-qPCR to confirm downstream gene regulation.
Main Results:
- PMFBP1 interacts with chaperonin-containing TCP1 subunit 3 (CCT3) and class I histone deacetylases, including HDAC3.
- Loss of PMFBP1 in mice leads to reduced HDAC levels and altered testicular proteomic profiles, affecting spermatogenesis and flagella assembly.
- Downstream response factors RNF151 and RNF133 are regulated by the PMFBP1-HDAC3 axis.
Conclusions:
- PMFBP1 interacts with CCT3, modulating HDAC3 expression, which subsequently downregulates RNF151 and RNF133, causing abnormal sperm phenotypes.
- This study uncovers a novel molecular mechanism for PMFBP1 in spermatogenesis.
- The findings highlight the utility of multi-omics approaches for gene functional annotation.
