Cargando…
Mechanism of human Lig1 regulation by PCNA in Okazaki fragment sealing
During lagging strand synthesis, DNA Ligase 1 (Lig1) cooperates with the sliding clamp PCNA to seal the nicks between Okazaki fragments generated by Pol δ and Flap endonuclease 1 (FEN1). We present several cryo-EM structures combined with functional assays, showing that human Lig1 recruits PCNA to n...
Autores principales: | , , , , , , , , |
---|---|
Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Nature Publishing Group UK
2022
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9767926/ https://www.ncbi.nlm.nih.gov/pubmed/36539424 http://dx.doi.org/10.1038/s41467-022-35475-z |
_version_ | 1784854061053378560 |
---|---|
author | Blair, Kerry Tehseen, Muhammad Raducanu, Vlad-Stefan Shahid, Taha Lancey, Claudia Rashid, Fahad Crehuet, Ramon Hamdan, Samir M. De Biasio, Alfredo |
author_facet | Blair, Kerry Tehseen, Muhammad Raducanu, Vlad-Stefan Shahid, Taha Lancey, Claudia Rashid, Fahad Crehuet, Ramon Hamdan, Samir M. De Biasio, Alfredo |
author_sort | Blair, Kerry |
collection | PubMed |
description | During lagging strand synthesis, DNA Ligase 1 (Lig1) cooperates with the sliding clamp PCNA to seal the nicks between Okazaki fragments generated by Pol δ and Flap endonuclease 1 (FEN1). We present several cryo-EM structures combined with functional assays, showing that human Lig1 recruits PCNA to nicked DNA using two PCNA-interacting motifs (PIPs) located at its disordered N-terminus (PIP(N-term)) and DNA binding domain (PIP(DBD)). Once Lig1 and PCNA assemble as two-stack rings encircling DNA, PIP(N-term) is released from PCNA and only PIP(DBD) is required for ligation to facilitate the substrate handoff from FEN1. Consistently, we observed that PCNA forms a defined complex with FEN1 and nicked DNA, and it recruits Lig1 to an unoccupied monomer creating a toolbelt that drives the transfer of DNA to Lig1. Collectively, our results provide a structural model on how PCNA regulates FEN1 and Lig1 during Okazaki fragments maturation. |
format | Online Article Text |
id | pubmed-9767926 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | Nature Publishing Group UK |
record_format | MEDLINE/PubMed |
spelling | pubmed-97679262022-12-22 Mechanism of human Lig1 regulation by PCNA in Okazaki fragment sealing Blair, Kerry Tehseen, Muhammad Raducanu, Vlad-Stefan Shahid, Taha Lancey, Claudia Rashid, Fahad Crehuet, Ramon Hamdan, Samir M. De Biasio, Alfredo Nat Commun Article During lagging strand synthesis, DNA Ligase 1 (Lig1) cooperates with the sliding clamp PCNA to seal the nicks between Okazaki fragments generated by Pol δ and Flap endonuclease 1 (FEN1). We present several cryo-EM structures combined with functional assays, showing that human Lig1 recruits PCNA to nicked DNA using two PCNA-interacting motifs (PIPs) located at its disordered N-terminus (PIP(N-term)) and DNA binding domain (PIP(DBD)). Once Lig1 and PCNA assemble as two-stack rings encircling DNA, PIP(N-term) is released from PCNA and only PIP(DBD) is required for ligation to facilitate the substrate handoff from FEN1. Consistently, we observed that PCNA forms a defined complex with FEN1 and nicked DNA, and it recruits Lig1 to an unoccupied monomer creating a toolbelt that drives the transfer of DNA to Lig1. Collectively, our results provide a structural model on how PCNA regulates FEN1 and Lig1 during Okazaki fragments maturation. Nature Publishing Group UK 2022-12-20 /pmc/articles/PMC9767926/ /pubmed/36539424 http://dx.doi.org/10.1038/s41467-022-35475-z Text en © The Author(s) 2022, corrected publication 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 Blair, Kerry Tehseen, Muhammad Raducanu, Vlad-Stefan Shahid, Taha Lancey, Claudia Rashid, Fahad Crehuet, Ramon Hamdan, Samir M. De Biasio, Alfredo Mechanism of human Lig1 regulation by PCNA in Okazaki fragment sealing |
title | Mechanism of human Lig1 regulation by PCNA in Okazaki fragment sealing |
title_full | Mechanism of human Lig1 regulation by PCNA in Okazaki fragment sealing |
title_fullStr | Mechanism of human Lig1 regulation by PCNA in Okazaki fragment sealing |
title_full_unstemmed | Mechanism of human Lig1 regulation by PCNA in Okazaki fragment sealing |
title_short | Mechanism of human Lig1 regulation by PCNA in Okazaki fragment sealing |
title_sort | mechanism of human lig1 regulation by pcna in okazaki fragment sealing |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9767926/ https://www.ncbi.nlm.nih.gov/pubmed/36539424 http://dx.doi.org/10.1038/s41467-022-35475-z |
work_keys_str_mv | AT blairkerry mechanismofhumanlig1regulationbypcnainokazakifragmentsealing AT tehseenmuhammad mechanismofhumanlig1regulationbypcnainokazakifragmentsealing AT raducanuvladstefan mechanismofhumanlig1regulationbypcnainokazakifragmentsealing AT shahidtaha mechanismofhumanlig1regulationbypcnainokazakifragmentsealing AT lanceyclaudia mechanismofhumanlig1regulationbypcnainokazakifragmentsealing AT rashidfahad mechanismofhumanlig1regulationbypcnainokazakifragmentsealing AT crehuetramon mechanismofhumanlig1regulationbypcnainokazakifragmentsealing AT hamdansamirm mechanismofhumanlig1regulationbypcnainokazakifragmentsealing AT debiasioalfredo mechanismofhumanlig1regulationbypcnainokazakifragmentsealing |