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Proximity-dependent biotin labeling in testicular germ cells identified TESMIN-associated proteins

Characterization of protein–protein interactions (PPI) is a key to understanding the functions of proteins of interest. Recently developed proximity-dependent biotin identification (BioID) has been actively investigated as an alternative PPI mapping method because of its usefulness in uncovering tra...

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Autores principales: Oura, Seiya, Ninomiya, Akinori, Sugihara, Fuminori, Matzuk, Martin M., Ikawa, Masahito
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group UK 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9789103/
https://www.ncbi.nlm.nih.gov/pubmed/36564444
http://dx.doi.org/10.1038/s41598-022-26501-7
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author Oura, Seiya
Ninomiya, Akinori
Sugihara, Fuminori
Matzuk, Martin M.
Ikawa, Masahito
author_facet Oura, Seiya
Ninomiya, Akinori
Sugihara, Fuminori
Matzuk, Martin M.
Ikawa, Masahito
author_sort Oura, Seiya
collection PubMed
description Characterization of protein–protein interactions (PPI) is a key to understanding the functions of proteins of interest. Recently developed proximity-dependent biotin identification (BioID) has been actively investigated as an alternative PPI mapping method because of its usefulness in uncovering transient PPI. Here, as an example of proximity labeling proteomics application in the testis, we generated two transgenic mouse lines expressing two biotin ligases (BioID2 or TurboID) fused with TESMIN, which translocates from the cytosol to the nucleus during meiotic progression and is required for reproduction. The BioID2 transgene, albeit not the TurboID transgene, rescued fertility defects of the Tesmin KO male mice, indicating that the TESMIN-BioID2 fusion can physiologically replace TESMIN. Furthermore, biotinylated protein pull-down and affinity-purification followed by mass spectrometry using the TESMIN-BioID2 transgenic mice captured components of the MYBL1–MuvB complex that regulate cell-cycle gene expression. Thus, our study shows that proximity labeling proteomics can be applied in male germ cells, although the choice of biotin ligase needs to be carefully tested.
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spelling pubmed-97891032022-12-25 Proximity-dependent biotin labeling in testicular germ cells identified TESMIN-associated proteins Oura, Seiya Ninomiya, Akinori Sugihara, Fuminori Matzuk, Martin M. Ikawa, Masahito Sci Rep Article Characterization of protein–protein interactions (PPI) is a key to understanding the functions of proteins of interest. Recently developed proximity-dependent biotin identification (BioID) has been actively investigated as an alternative PPI mapping method because of its usefulness in uncovering transient PPI. Here, as an example of proximity labeling proteomics application in the testis, we generated two transgenic mouse lines expressing two biotin ligases (BioID2 or TurboID) fused with TESMIN, which translocates from the cytosol to the nucleus during meiotic progression and is required for reproduction. The BioID2 transgene, albeit not the TurboID transgene, rescued fertility defects of the Tesmin KO male mice, indicating that the TESMIN-BioID2 fusion can physiologically replace TESMIN. Furthermore, biotinylated protein pull-down and affinity-purification followed by mass spectrometry using the TESMIN-BioID2 transgenic mice captured components of the MYBL1–MuvB complex that regulate cell-cycle gene expression. Thus, our study shows that proximity labeling proteomics can be applied in male germ cells, although the choice of biotin ligase needs to be carefully tested. Nature Publishing Group UK 2022-12-23 /pmc/articles/PMC9789103/ /pubmed/36564444 http://dx.doi.org/10.1038/s41598-022-26501-7 Text en © The Author(s) 2022 https://creativecommons.org/licenses/by/4.0/Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) .
spellingShingle Article
Oura, Seiya
Ninomiya, Akinori
Sugihara, Fuminori
Matzuk, Martin M.
Ikawa, Masahito
Proximity-dependent biotin labeling in testicular germ cells identified TESMIN-associated proteins
title Proximity-dependent biotin labeling in testicular germ cells identified TESMIN-associated proteins
title_full Proximity-dependent biotin labeling in testicular germ cells identified TESMIN-associated proteins
title_fullStr Proximity-dependent biotin labeling in testicular germ cells identified TESMIN-associated proteins
title_full_unstemmed Proximity-dependent biotin labeling in testicular germ cells identified TESMIN-associated proteins
title_short Proximity-dependent biotin labeling in testicular germ cells identified TESMIN-associated proteins
title_sort proximity-dependent biotin labeling in testicular germ cells identified tesmin-associated proteins
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9789103/
https://www.ncbi.nlm.nih.gov/pubmed/36564444
http://dx.doi.org/10.1038/s41598-022-26501-7
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