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Proliferating Cell Nuclear Antigen (PCNA) Interactions in Solution Studied by NMR

PCNA is an essential factor for DNA replication and repair. It forms a ring shaped structure of 86 kDa by the symmetric association of three identical protomers. The ring encircles the DNA and acts as a docking platform for other proteins, most of them containing the PCNA Interaction Protein sequenc...

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Detalles Bibliográficos
Autores principales: De Biasio, Alfredo, Campos-Olivas, Ramón, Sánchez, Ricardo, López-Alonso, Jorge P., Pantoja-Uceda, David, Merino, Nekane, Villate, Maider, Martin-Garcia, Jose M., Castillo, Francisco, Luque, Irene, Blanco, Francisco J.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2012
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3491057/
https://www.ncbi.nlm.nih.gov/pubmed/23139781
http://dx.doi.org/10.1371/journal.pone.0048390
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author De Biasio, Alfredo
Campos-Olivas, Ramón
Sánchez, Ricardo
López-Alonso, Jorge P.
Pantoja-Uceda, David
Merino, Nekane
Villate, Maider
Martin-Garcia, Jose M.
Castillo, Francisco
Luque, Irene
Blanco, Francisco J.
author_facet De Biasio, Alfredo
Campos-Olivas, Ramón
Sánchez, Ricardo
López-Alonso, Jorge P.
Pantoja-Uceda, David
Merino, Nekane
Villate, Maider
Martin-Garcia, Jose M.
Castillo, Francisco
Luque, Irene
Blanco, Francisco J.
author_sort De Biasio, Alfredo
collection PubMed
description PCNA is an essential factor for DNA replication and repair. It forms a ring shaped structure of 86 kDa by the symmetric association of three identical protomers. The ring encircles the DNA and acts as a docking platform for other proteins, most of them containing the PCNA Interaction Protein sequence (PIP-box). We have used NMR to characterize the interactions of PCNA with several other proteins and fragments in solution. The binding of the PIP-box peptide of the cell cycle inhibitor p21 to PCNA is consistent with the crystal structure of the complex. A shorter p21 peptide binds with reduced affinity but retains most of the molecular recognition determinants. However the binding of the corresponding peptide of the tumor suppressor ING1 is extremely weak, indicating that slight deviations from the consensus PIP-box sequence dramatically reduce the affinity for PCNA, in contrast with a proposed less stringent PIP-box sequence requirement. We could not detect any binding between PCNA and the MCL-1 or the CDK2 protein, reported to interact with PCNA in biochemical assays. This suggests that they do not bind directly to PCNA, or they do but very weakly, with additional unidentified factors stabilizing the interactions in the cell. Backbone dynamics measurements show three PCNA regions with high relative flexibility, including the interdomain connector loop (IDCL) and the C-terminus, both of them involved in the interaction with the PIP-box. Our work provides the basis for high resolution studies of direct ligand binding to PCNA in solution.
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spelling pubmed-34910572012-11-08 Proliferating Cell Nuclear Antigen (PCNA) Interactions in Solution Studied by NMR De Biasio, Alfredo Campos-Olivas, Ramón Sánchez, Ricardo López-Alonso, Jorge P. Pantoja-Uceda, David Merino, Nekane Villate, Maider Martin-Garcia, Jose M. Castillo, Francisco Luque, Irene Blanco, Francisco J. PLoS One Research Article PCNA is an essential factor for DNA replication and repair. It forms a ring shaped structure of 86 kDa by the symmetric association of three identical protomers. The ring encircles the DNA and acts as a docking platform for other proteins, most of them containing the PCNA Interaction Protein sequence (PIP-box). We have used NMR to characterize the interactions of PCNA with several other proteins and fragments in solution. The binding of the PIP-box peptide of the cell cycle inhibitor p21 to PCNA is consistent with the crystal structure of the complex. A shorter p21 peptide binds with reduced affinity but retains most of the molecular recognition determinants. However the binding of the corresponding peptide of the tumor suppressor ING1 is extremely weak, indicating that slight deviations from the consensus PIP-box sequence dramatically reduce the affinity for PCNA, in contrast with a proposed less stringent PIP-box sequence requirement. We could not detect any binding between PCNA and the MCL-1 or the CDK2 protein, reported to interact with PCNA in biochemical assays. This suggests that they do not bind directly to PCNA, or they do but very weakly, with additional unidentified factors stabilizing the interactions in the cell. Backbone dynamics measurements show three PCNA regions with high relative flexibility, including the interdomain connector loop (IDCL) and the C-terminus, both of them involved in the interaction with the PIP-box. Our work provides the basis for high resolution studies of direct ligand binding to PCNA in solution. Public Library of Science 2012-11-06 /pmc/articles/PMC3491057/ /pubmed/23139781 http://dx.doi.org/10.1371/journal.pone.0048390 Text en © 2012 De Biasio et al http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.
spellingShingle Research Article
De Biasio, Alfredo
Campos-Olivas, Ramón
Sánchez, Ricardo
López-Alonso, Jorge P.
Pantoja-Uceda, David
Merino, Nekane
Villate, Maider
Martin-Garcia, Jose M.
Castillo, Francisco
Luque, Irene
Blanco, Francisco J.
Proliferating Cell Nuclear Antigen (PCNA) Interactions in Solution Studied by NMR
title Proliferating Cell Nuclear Antigen (PCNA) Interactions in Solution Studied by NMR
title_full Proliferating Cell Nuclear Antigen (PCNA) Interactions in Solution Studied by NMR
title_fullStr Proliferating Cell Nuclear Antigen (PCNA) Interactions in Solution Studied by NMR
title_full_unstemmed Proliferating Cell Nuclear Antigen (PCNA) Interactions in Solution Studied by NMR
title_short Proliferating Cell Nuclear Antigen (PCNA) Interactions in Solution Studied by NMR
title_sort proliferating cell nuclear antigen (pcna) interactions in solution studied by nmr
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3491057/
https://www.ncbi.nlm.nih.gov/pubmed/23139781
http://dx.doi.org/10.1371/journal.pone.0048390
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