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The unifying catalytic mechanism of the RING-between-RING E3 ubiquitin ligase family

The RING-between-RING (RBR) E3 ubiquitin ligase family in humans comprises 14 members and is defined by a two-step catalytic mechanism in which ubiquitin is first transferred from an E2 ubiquitin-conjugating enzyme to the RBR active site and then to the substrate. To define the core features of this...

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Autores principales: Wang, Xiangyi S., Cotton, Thomas R., Trevelyan, Sarah J., Richardson, Lachlan W., Lee, Wei Ting, Silke, John, Lechtenberg, Bernhard C.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group UK 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9834252/
https://www.ncbi.nlm.nih.gov/pubmed/36631489
http://dx.doi.org/10.1038/s41467-023-35871-z
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author Wang, Xiangyi S.
Cotton, Thomas R.
Trevelyan, Sarah J.
Richardson, Lachlan W.
Lee, Wei Ting
Silke, John
Lechtenberg, Bernhard C.
author_facet Wang, Xiangyi S.
Cotton, Thomas R.
Trevelyan, Sarah J.
Richardson, Lachlan W.
Lee, Wei Ting
Silke, John
Lechtenberg, Bernhard C.
author_sort Wang, Xiangyi S.
collection PubMed
description The RING-between-RING (RBR) E3 ubiquitin ligase family in humans comprises 14 members and is defined by a two-step catalytic mechanism in which ubiquitin is first transferred from an E2 ubiquitin-conjugating enzyme to the RBR active site and then to the substrate. To define the core features of this catalytic mechanism, we here structurally and biochemically characterise the two RBRs HOIL-1 and RNF216. Crystal structures of both enzymes in their RBR/E2-Ub/Ub transthiolation complexes capturing the first catalytic step, together with complementary functional experiments, reveal the defining features of the RBR catalytic mechanism. RBRs catalyse ubiquitination via a conserved transthiolation complex structure that enables efficient E2-to-RBR ubiquitin transfer. Our data also highlight a conserved RBR allosteric activation mechanism by distinct ubiquitin linkages that suggests RBRs employ a feed-forward mechanism. We finally identify that the HOIL-1 RING2 domain contains an unusual Zn2/Cys6 binuclear cluster that is required for catalytic activity and substrate ubiquitination.
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spelling pubmed-98342522023-01-13 The unifying catalytic mechanism of the RING-between-RING E3 ubiquitin ligase family Wang, Xiangyi S. Cotton, Thomas R. Trevelyan, Sarah J. Richardson, Lachlan W. Lee, Wei Ting Silke, John Lechtenberg, Bernhard C. Nat Commun Article The RING-between-RING (RBR) E3 ubiquitin ligase family in humans comprises 14 members and is defined by a two-step catalytic mechanism in which ubiquitin is first transferred from an E2 ubiquitin-conjugating enzyme to the RBR active site and then to the substrate. To define the core features of this catalytic mechanism, we here structurally and biochemically characterise the two RBRs HOIL-1 and RNF216. Crystal structures of both enzymes in their RBR/E2-Ub/Ub transthiolation complexes capturing the first catalytic step, together with complementary functional experiments, reveal the defining features of the RBR catalytic mechanism. RBRs catalyse ubiquitination via a conserved transthiolation complex structure that enables efficient E2-to-RBR ubiquitin transfer. Our data also highlight a conserved RBR allosteric activation mechanism by distinct ubiquitin linkages that suggests RBRs employ a feed-forward mechanism. We finally identify that the HOIL-1 RING2 domain contains an unusual Zn2/Cys6 binuclear cluster that is required for catalytic activity and substrate ubiquitination. Nature Publishing Group UK 2023-01-11 /pmc/articles/PMC9834252/ /pubmed/36631489 http://dx.doi.org/10.1038/s41467-023-35871-z Text en © The Author(s) 2023 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
Wang, Xiangyi S.
Cotton, Thomas R.
Trevelyan, Sarah J.
Richardson, Lachlan W.
Lee, Wei Ting
Silke, John
Lechtenberg, Bernhard C.
The unifying catalytic mechanism of the RING-between-RING E3 ubiquitin ligase family
title The unifying catalytic mechanism of the RING-between-RING E3 ubiquitin ligase family
title_full The unifying catalytic mechanism of the RING-between-RING E3 ubiquitin ligase family
title_fullStr The unifying catalytic mechanism of the RING-between-RING E3 ubiquitin ligase family
title_full_unstemmed The unifying catalytic mechanism of the RING-between-RING E3 ubiquitin ligase family
title_short The unifying catalytic mechanism of the RING-between-RING E3 ubiquitin ligase family
title_sort unifying catalytic mechanism of the ring-between-ring e3 ubiquitin ligase family
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9834252/
https://www.ncbi.nlm.nih.gov/pubmed/36631489
http://dx.doi.org/10.1038/s41467-023-35871-z
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