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Molecular basis of ubiquitin-specific protease 8 autoinhibition by the WW-like domain
Ubiquitin-specific protease 8 (USP8) is a deubiquitinating enzyme involved in multiple membrane trafficking pathways. The enzyme activity is inhibited by binding to 14-3-3 proteins. Mutations in the 14-3-3-binding motif in USP8 are related to Cushing’s disease. However, the molecular basis of USP8 a...
Autores principales: | , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
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Nature Publishing Group UK
2021
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8576004/ https://www.ncbi.nlm.nih.gov/pubmed/34750505 http://dx.doi.org/10.1038/s42003-021-02802-x |
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author | Kakihara, Keijun Asamizu, Kengo Moritsugu, Kei Kubo, Masahide Kitaguchi, Tetsuya Endo, Akinori Kidera, Akinori Ikeguchi, Mitsunori Kato, Akira Komada, Masayuki Fukushima, Toshiaki |
author_facet | Kakihara, Keijun Asamizu, Kengo Moritsugu, Kei Kubo, Masahide Kitaguchi, Tetsuya Endo, Akinori Kidera, Akinori Ikeguchi, Mitsunori Kato, Akira Komada, Masayuki Fukushima, Toshiaki |
author_sort | Kakihara, Keijun |
collection | PubMed |
description | Ubiquitin-specific protease 8 (USP8) is a deubiquitinating enzyme involved in multiple membrane trafficking pathways. The enzyme activity is inhibited by binding to 14-3-3 proteins. Mutations in the 14-3-3-binding motif in USP8 are related to Cushing’s disease. However, the molecular basis of USP8 activity regulation remains unclear. This study identified amino acids 645–684 of USP8 as an autoinhibitory region, which might interact with the catalytic USP domain, as per the results of pull-down and single-molecule FRET assays performed in this study. In silico modelling indicated that the region forms a WW-like domain structure, plugs the catalytic cleft, and narrows the entrance to the ubiquitin-binding pocket. Furthermore, 14-3-3 inhibited USP8 activity partly by enhancing the interaction between the WW-like and USP domains. These findings provide the molecular basis of USP8 autoinhibition via the WW-like domain. Moreover, they suggest that the release of autoinhibition may underlie Cushing’s disease due to USP8 mutations. |
format | Online Article Text |
id | pubmed-8576004 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2021 |
publisher | Nature Publishing Group UK |
record_format | MEDLINE/PubMed |
spelling | pubmed-85760042021-11-19 Molecular basis of ubiquitin-specific protease 8 autoinhibition by the WW-like domain Kakihara, Keijun Asamizu, Kengo Moritsugu, Kei Kubo, Masahide Kitaguchi, Tetsuya Endo, Akinori Kidera, Akinori Ikeguchi, Mitsunori Kato, Akira Komada, Masayuki Fukushima, Toshiaki Commun Biol Article Ubiquitin-specific protease 8 (USP8) is a deubiquitinating enzyme involved in multiple membrane trafficking pathways. The enzyme activity is inhibited by binding to 14-3-3 proteins. Mutations in the 14-3-3-binding motif in USP8 are related to Cushing’s disease. However, the molecular basis of USP8 activity regulation remains unclear. This study identified amino acids 645–684 of USP8 as an autoinhibitory region, which might interact with the catalytic USP domain, as per the results of pull-down and single-molecule FRET assays performed in this study. In silico modelling indicated that the region forms a WW-like domain structure, plugs the catalytic cleft, and narrows the entrance to the ubiquitin-binding pocket. Furthermore, 14-3-3 inhibited USP8 activity partly by enhancing the interaction between the WW-like and USP domains. These findings provide the molecular basis of USP8 autoinhibition via the WW-like domain. Moreover, they suggest that the release of autoinhibition may underlie Cushing’s disease due to USP8 mutations. Nature Publishing Group UK 2021-11-08 /pmc/articles/PMC8576004/ /pubmed/34750505 http://dx.doi.org/10.1038/s42003-021-02802-x Text en © The Author(s) 2021 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 license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license 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 license, visit http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) . |
spellingShingle | Article Kakihara, Keijun Asamizu, Kengo Moritsugu, Kei Kubo, Masahide Kitaguchi, Tetsuya Endo, Akinori Kidera, Akinori Ikeguchi, Mitsunori Kato, Akira Komada, Masayuki Fukushima, Toshiaki Molecular basis of ubiquitin-specific protease 8 autoinhibition by the WW-like domain |
title | Molecular basis of ubiquitin-specific protease 8 autoinhibition by the WW-like domain |
title_full | Molecular basis of ubiquitin-specific protease 8 autoinhibition by the WW-like domain |
title_fullStr | Molecular basis of ubiquitin-specific protease 8 autoinhibition by the WW-like domain |
title_full_unstemmed | Molecular basis of ubiquitin-specific protease 8 autoinhibition by the WW-like domain |
title_short | Molecular basis of ubiquitin-specific protease 8 autoinhibition by the WW-like domain |
title_sort | molecular basis of ubiquitin-specific protease 8 autoinhibition by the ww-like domain |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8576004/ https://www.ncbi.nlm.nih.gov/pubmed/34750505 http://dx.doi.org/10.1038/s42003-021-02802-x |
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