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The role of SLX4 and its associated nucleases in DNA interstrand crosslink repair

A key step in the Fanconi anemia pathway of DNA interstrand crosslink (ICL) repair is the ICL unhooking by dual endonucleolytic incisions. SLX4/FANCP is a large scaffold protein that plays a central role in ICL unhooking. It contains multiple domains that interact with many proteins including three...

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Autores principales: Hoogenboom, Wouter S, Boonen, Rick A C M, Knipscheer, Puck
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Oxford University Press 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6411836/
https://www.ncbi.nlm.nih.gov/pubmed/30576517
http://dx.doi.org/10.1093/nar/gky1276
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author Hoogenboom, Wouter S
Boonen, Rick A C M
Knipscheer, Puck
author_facet Hoogenboom, Wouter S
Boonen, Rick A C M
Knipscheer, Puck
author_sort Hoogenboom, Wouter S
collection PubMed
description A key step in the Fanconi anemia pathway of DNA interstrand crosslink (ICL) repair is the ICL unhooking by dual endonucleolytic incisions. SLX4/FANCP is a large scaffold protein that plays a central role in ICL unhooking. It contains multiple domains that interact with many proteins including three different endonucleases and also acts in several other DNA repair pathways. While it is known that its interaction with the endonuclease XPF-ERCC1 is required for its function in ICL repair, which other domains act in this process is unclear. Here, we used Xenopus egg extracts to determine ICL repair specific features of SLX4. We show that the SLX4-interacting endonuclease SLX1 is not required for ICL repair and demonstrate that all essential SLX4 domains are located at the N-terminal half of the protein. The MLR domain is crucial for the recruitment of XPF-ERCC1 but also has an unanticipated function in recruiting SLX4 to the site of damage. Although we find the BTB is not essential for ICL repair in our system, dimerization of SLX4 could be important. Our data provide new insights into the mechanism by which SLX4 acts in ICL repair.
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spelling pubmed-64118362019-03-15 The role of SLX4 and its associated nucleases in DNA interstrand crosslink repair Hoogenboom, Wouter S Boonen, Rick A C M Knipscheer, Puck Nucleic Acids Res Genome Integrity, Repair and Replication A key step in the Fanconi anemia pathway of DNA interstrand crosslink (ICL) repair is the ICL unhooking by dual endonucleolytic incisions. SLX4/FANCP is a large scaffold protein that plays a central role in ICL unhooking. It contains multiple domains that interact with many proteins including three different endonucleases and also acts in several other DNA repair pathways. While it is known that its interaction with the endonuclease XPF-ERCC1 is required for its function in ICL repair, which other domains act in this process is unclear. Here, we used Xenopus egg extracts to determine ICL repair specific features of SLX4. We show that the SLX4-interacting endonuclease SLX1 is not required for ICL repair and demonstrate that all essential SLX4 domains are located at the N-terminal half of the protein. The MLR domain is crucial for the recruitment of XPF-ERCC1 but also has an unanticipated function in recruiting SLX4 to the site of damage. Although we find the BTB is not essential for ICL repair in our system, dimerization of SLX4 could be important. Our data provide new insights into the mechanism by which SLX4 acts in ICL repair. Oxford University Press 2019-03-18 2018-12-21 /pmc/articles/PMC6411836/ /pubmed/30576517 http://dx.doi.org/10.1093/nar/gky1276 Text en © The Author(s) 2018. Published by Oxford University Press on behalf of Nucleic Acids Research. http://creativecommons.org/licenses/by-nc/4.0/ This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/4.0/), which permits non-commercial re-use, distribution, and reproduction in any medium, provided the original work is properly cited. For commercial re-use, please contact journals.permissions@oup.com
spellingShingle Genome Integrity, Repair and Replication
Hoogenboom, Wouter S
Boonen, Rick A C M
Knipscheer, Puck
The role of SLX4 and its associated nucleases in DNA interstrand crosslink repair
title The role of SLX4 and its associated nucleases in DNA interstrand crosslink repair
title_full The role of SLX4 and its associated nucleases in DNA interstrand crosslink repair
title_fullStr The role of SLX4 and its associated nucleases in DNA interstrand crosslink repair
title_full_unstemmed The role of SLX4 and its associated nucleases in DNA interstrand crosslink repair
title_short The role of SLX4 and its associated nucleases in DNA interstrand crosslink repair
title_sort role of slx4 and its associated nucleases in dna interstrand crosslink repair
topic Genome Integrity, Repair and Replication
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6411836/
https://www.ncbi.nlm.nih.gov/pubmed/30576517
http://dx.doi.org/10.1093/nar/gky1276
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