Cargando…
Role of Nse1 Subunit of SMC5/6 Complex as a Ubiquitin Ligase
Structural Maintenance of Chromosomes (SMC) complexes are important for many aspects of the chromosomal organization. Unlike cohesin and condensin, the SMC5/6 complex contains a variant RING domain carried by its Nse1 subunit. RING domains are characteristic for ubiquitin ligases, and human NSE1 has...
Autores principales: | , , , , |
---|---|
Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
MDPI
2022
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8750328/ https://www.ncbi.nlm.nih.gov/pubmed/35011726 http://dx.doi.org/10.3390/cells11010165 |
_version_ | 1784631435302273024 |
---|---|
author | Kolesar, Peter Stejskal, Karel Potesil, David Murray, Johanne M. Palecek, Jan J. |
author_facet | Kolesar, Peter Stejskal, Karel Potesil, David Murray, Johanne M. Palecek, Jan J. |
author_sort | Kolesar, Peter |
collection | PubMed |
description | Structural Maintenance of Chromosomes (SMC) complexes are important for many aspects of the chromosomal organization. Unlike cohesin and condensin, the SMC5/6 complex contains a variant RING domain carried by its Nse1 subunit. RING domains are characteristic for ubiquitin ligases, and human NSE1 has been shown to possess ubiquitin-ligase activity in vitro. However, other studies were unable to show such activity. Here, we confirm Nse1 ubiquitin-ligase activity using purified Schizosaccharomyces pombe proteins. We demonstrate that the Nse1 ligase activity is stimulated by Nse3 and Nse4. We show that Nse1 specifically utilizes Ubc13/Mms2 E2 enzyme and interacts directly with ubiquitin. We identify the Nse1 mutation (R188E) that specifically disrupts its E3 activity and demonstrate that the Nse1-dependent ubiquitination is particularly important under replication stress. Moreover, we determine Nse4 (lysine K181) as the first known SMC5/6-associated Nse1 substrate. Interestingly, abolition of Nse4 modification at K181 leads to suppression of DNA-damage sensitivity of other SMC5/6 mutants. Altogether, this study brings new evidence for Nse1 ubiquitin ligase activity, significantly advancing our understanding of this enigmatic SMC5/6 function. |
format | Online Article Text |
id | pubmed-8750328 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | MDPI |
record_format | MEDLINE/PubMed |
spelling | pubmed-87503282022-01-12 Role of Nse1 Subunit of SMC5/6 Complex as a Ubiquitin Ligase Kolesar, Peter Stejskal, Karel Potesil, David Murray, Johanne M. Palecek, Jan J. Cells Article Structural Maintenance of Chromosomes (SMC) complexes are important for many aspects of the chromosomal organization. Unlike cohesin and condensin, the SMC5/6 complex contains a variant RING domain carried by its Nse1 subunit. RING domains are characteristic for ubiquitin ligases, and human NSE1 has been shown to possess ubiquitin-ligase activity in vitro. However, other studies were unable to show such activity. Here, we confirm Nse1 ubiquitin-ligase activity using purified Schizosaccharomyces pombe proteins. We demonstrate that the Nse1 ligase activity is stimulated by Nse3 and Nse4. We show that Nse1 specifically utilizes Ubc13/Mms2 E2 enzyme and interacts directly with ubiquitin. We identify the Nse1 mutation (R188E) that specifically disrupts its E3 activity and demonstrate that the Nse1-dependent ubiquitination is particularly important under replication stress. Moreover, we determine Nse4 (lysine K181) as the first known SMC5/6-associated Nse1 substrate. Interestingly, abolition of Nse4 modification at K181 leads to suppression of DNA-damage sensitivity of other SMC5/6 mutants. Altogether, this study brings new evidence for Nse1 ubiquitin ligase activity, significantly advancing our understanding of this enigmatic SMC5/6 function. MDPI 2022-01-04 /pmc/articles/PMC8750328/ /pubmed/35011726 http://dx.doi.org/10.3390/cells11010165 Text en © 2022 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/). |
spellingShingle | Article Kolesar, Peter Stejskal, Karel Potesil, David Murray, Johanne M. Palecek, Jan J. Role of Nse1 Subunit of SMC5/6 Complex as a Ubiquitin Ligase |
title | Role of Nse1 Subunit of SMC5/6 Complex as a Ubiquitin Ligase |
title_full | Role of Nse1 Subunit of SMC5/6 Complex as a Ubiquitin Ligase |
title_fullStr | Role of Nse1 Subunit of SMC5/6 Complex as a Ubiquitin Ligase |
title_full_unstemmed | Role of Nse1 Subunit of SMC5/6 Complex as a Ubiquitin Ligase |
title_short | Role of Nse1 Subunit of SMC5/6 Complex as a Ubiquitin Ligase |
title_sort | role of nse1 subunit of smc5/6 complex as a ubiquitin ligase |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8750328/ https://www.ncbi.nlm.nih.gov/pubmed/35011726 http://dx.doi.org/10.3390/cells11010165 |
work_keys_str_mv | AT kolesarpeter roleofnse1subunitofsmc56complexasaubiquitinligase AT stejskalkarel roleofnse1subunitofsmc56complexasaubiquitinligase AT potesildavid roleofnse1subunitofsmc56complexasaubiquitinligase AT murrayjohannem roleofnse1subunitofsmc56complexasaubiquitinligase AT palecekjanj roleofnse1subunitofsmc56complexasaubiquitinligase |