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A unique arginine cluster in PolDIP2 enhances nucleotide binding and DNA synthesis by PrimPol
Replication forks often stall at damaged DNA. To overcome these obstructions and complete the DNA duplication in a timely fashion, replication can be restarted downstream of the DNA lesion. In mammalian cells, this repriming of replication can be achieved through the activities of primase and polyme...
Autores principales: | , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Oxford University Press
2021
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7913696/ https://www.ncbi.nlm.nih.gov/pubmed/33533925 http://dx.doi.org/10.1093/nar/gkab049 |
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author | Kasho, Kazutoshi Stojkovič, Gorazd Velázquez-Ruiz, Cristina Martínez-Jiménez, Maria Isabel Doimo, Mara Laurent, Timothée Berner, Andreas Pérez-Rivera, Aldo E Jenninger, Louise Blanco, Luis Wanrooij, Sjoerd |
author_facet | Kasho, Kazutoshi Stojkovič, Gorazd Velázquez-Ruiz, Cristina Martínez-Jiménez, Maria Isabel Doimo, Mara Laurent, Timothée Berner, Andreas Pérez-Rivera, Aldo E Jenninger, Louise Blanco, Luis Wanrooij, Sjoerd |
author_sort | Kasho, Kazutoshi |
collection | PubMed |
description | Replication forks often stall at damaged DNA. To overcome these obstructions and complete the DNA duplication in a timely fashion, replication can be restarted downstream of the DNA lesion. In mammalian cells, this repriming of replication can be achieved through the activities of primase and polymerase PrimPol. PrimPol is stimulated in DNA synthesis through interaction with PolDIP2, however the exact mechanism of this PolDIP2-dependent stimulation is still unclear. Here, we show that PrimPol uses a flexible loop to interact with the C-terminal ApaG-like domain of PolDIP2, and that this contact is essential for PrimPol's enhanced processivity. PolDIP2 increases primer-template and dNTP binding affinities of PrimPol, which concomitantly enhances its nucleotide incorporation efficiency. This stimulation is dependent on a unique arginine cluster in PolDIP2. Since the polymerase activity of PrimPol alone is very limited, this mechanism, where the affinity for dNTPs gets increased by PolDIP2 binding, might be critical for the in vivo function of PrimPol in tolerating DNA lesions at physiological nucleotide concentrations. |
format | Online Article Text |
id | pubmed-7913696 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2021 |
publisher | Oxford University Press |
record_format | MEDLINE/PubMed |
spelling | pubmed-79136962021-03-03 A unique arginine cluster in PolDIP2 enhances nucleotide binding and DNA synthesis by PrimPol Kasho, Kazutoshi Stojkovič, Gorazd Velázquez-Ruiz, Cristina Martínez-Jiménez, Maria Isabel Doimo, Mara Laurent, Timothée Berner, Andreas Pérez-Rivera, Aldo E Jenninger, Louise Blanco, Luis Wanrooij, Sjoerd Nucleic Acids Res Nucleic Acid Enzymes Replication forks often stall at damaged DNA. To overcome these obstructions and complete the DNA duplication in a timely fashion, replication can be restarted downstream of the DNA lesion. In mammalian cells, this repriming of replication can be achieved through the activities of primase and polymerase PrimPol. PrimPol is stimulated in DNA synthesis through interaction with PolDIP2, however the exact mechanism of this PolDIP2-dependent stimulation is still unclear. Here, we show that PrimPol uses a flexible loop to interact with the C-terminal ApaG-like domain of PolDIP2, and that this contact is essential for PrimPol's enhanced processivity. PolDIP2 increases primer-template and dNTP binding affinities of PrimPol, which concomitantly enhances its nucleotide incorporation efficiency. This stimulation is dependent on a unique arginine cluster in PolDIP2. Since the polymerase activity of PrimPol alone is very limited, this mechanism, where the affinity for dNTPs gets increased by PolDIP2 binding, might be critical for the in vivo function of PrimPol in tolerating DNA lesions at physiological nucleotide concentrations. Oxford University Press 2021-02-03 /pmc/articles/PMC7913696/ /pubmed/33533925 http://dx.doi.org/10.1093/nar/gkab049 Text en © The Author(s) 2021. Published by Oxford University Press on behalf of Nucleic Acids Research. http://creativecommons.org/licenses/by/4.0/ This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited. |
spellingShingle | Nucleic Acid Enzymes Kasho, Kazutoshi Stojkovič, Gorazd Velázquez-Ruiz, Cristina Martínez-Jiménez, Maria Isabel Doimo, Mara Laurent, Timothée Berner, Andreas Pérez-Rivera, Aldo E Jenninger, Louise Blanco, Luis Wanrooij, Sjoerd A unique arginine cluster in PolDIP2 enhances nucleotide binding and DNA synthesis by PrimPol |
title | A unique arginine cluster in PolDIP2 enhances nucleotide binding and DNA synthesis by PrimPol |
title_full | A unique arginine cluster in PolDIP2 enhances nucleotide binding and DNA synthesis by PrimPol |
title_fullStr | A unique arginine cluster in PolDIP2 enhances nucleotide binding and DNA synthesis by PrimPol |
title_full_unstemmed | A unique arginine cluster in PolDIP2 enhances nucleotide binding and DNA synthesis by PrimPol |
title_short | A unique arginine cluster in PolDIP2 enhances nucleotide binding and DNA synthesis by PrimPol |
title_sort | unique arginine cluster in poldip2 enhances nucleotide binding and dna synthesis by primpol |
topic | Nucleic Acid Enzymes |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7913696/ https://www.ncbi.nlm.nih.gov/pubmed/33533925 http://dx.doi.org/10.1093/nar/gkab049 |
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