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Escherichia coli DnaE Polymerase Couples Pyrophosphatase Activity to DNA Replication
DNA Polymerases generate pyrophosphate every time they catalyze a step of DNA elongation. This elongation reaction is generally believed as thermodynamically favoured by the hydrolysis of pyrophosphate, catalyzed by inorganic pyrophosphatases. However, the specific action of inorganic pyrophosphatas...
Autores principales: | , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Public Library of Science
2016
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4822814/ https://www.ncbi.nlm.nih.gov/pubmed/27050298 http://dx.doi.org/10.1371/journal.pone.0152915 |
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author | Lapenta, Fabio Montón Silva, Alejandro Brandimarti, Renato Lanzi, Massimiliano Gratani, Fabio Lino Vellosillo Gonzalez, Perceval Perticarari, Sofia Hochkoeppler, Alejandro |
author_facet | Lapenta, Fabio Montón Silva, Alejandro Brandimarti, Renato Lanzi, Massimiliano Gratani, Fabio Lino Vellosillo Gonzalez, Perceval Perticarari, Sofia Hochkoeppler, Alejandro |
author_sort | Lapenta, Fabio |
collection | PubMed |
description | DNA Polymerases generate pyrophosphate every time they catalyze a step of DNA elongation. This elongation reaction is generally believed as thermodynamically favoured by the hydrolysis of pyrophosphate, catalyzed by inorganic pyrophosphatases. However, the specific action of inorganic pyrophosphatases coupled to DNA replication in vivo was never demonstrated. Here we show that the Polymerase-Histidinol-Phosphatase (PHP) domain of Escherichia coli DNA Polymerase III α subunit features pyrophosphatase activity. We also show that this activity is inhibited by fluoride, as commonly observed for inorganic pyrophosphatases, and we identified 3 amino acids of the PHP active site. Remarkably, E. coli cells expressing variants of these catalytic residues of α subunit feature aberrant phenotypes, poor viability, and are subject to high mutation frequencies. Our findings indicate that DNA Polymerases can couple DNA elongation and pyrophosphate hydrolysis, providing a mechanism for the control of DNA extension rate, and suggest a promising target for novel antibiotics. |
format | Online Article Text |
id | pubmed-4822814 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2016 |
publisher | Public Library of Science |
record_format | MEDLINE/PubMed |
spelling | pubmed-48228142016-04-22 Escherichia coli DnaE Polymerase Couples Pyrophosphatase Activity to DNA Replication Lapenta, Fabio Montón Silva, Alejandro Brandimarti, Renato Lanzi, Massimiliano Gratani, Fabio Lino Vellosillo Gonzalez, Perceval Perticarari, Sofia Hochkoeppler, Alejandro PLoS One Research Article DNA Polymerases generate pyrophosphate every time they catalyze a step of DNA elongation. This elongation reaction is generally believed as thermodynamically favoured by the hydrolysis of pyrophosphate, catalyzed by inorganic pyrophosphatases. However, the specific action of inorganic pyrophosphatases coupled to DNA replication in vivo was never demonstrated. Here we show that the Polymerase-Histidinol-Phosphatase (PHP) domain of Escherichia coli DNA Polymerase III α subunit features pyrophosphatase activity. We also show that this activity is inhibited by fluoride, as commonly observed for inorganic pyrophosphatases, and we identified 3 amino acids of the PHP active site. Remarkably, E. coli cells expressing variants of these catalytic residues of α subunit feature aberrant phenotypes, poor viability, and are subject to high mutation frequencies. Our findings indicate that DNA Polymerases can couple DNA elongation and pyrophosphate hydrolysis, providing a mechanism for the control of DNA extension rate, and suggest a promising target for novel antibiotics. Public Library of Science 2016-04-06 /pmc/articles/PMC4822814/ /pubmed/27050298 http://dx.doi.org/10.1371/journal.pone.0152915 Text en © 2016 Lapenta 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 (http://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. |
spellingShingle | Research Article Lapenta, Fabio Montón Silva, Alejandro Brandimarti, Renato Lanzi, Massimiliano Gratani, Fabio Lino Vellosillo Gonzalez, Perceval Perticarari, Sofia Hochkoeppler, Alejandro Escherichia coli DnaE Polymerase Couples Pyrophosphatase Activity to DNA Replication |
title | Escherichia coli DnaE Polymerase Couples Pyrophosphatase Activity to DNA Replication |
title_full | Escherichia coli DnaE Polymerase Couples Pyrophosphatase Activity to DNA Replication |
title_fullStr | Escherichia coli DnaE Polymerase Couples Pyrophosphatase Activity to DNA Replication |
title_full_unstemmed | Escherichia coli DnaE Polymerase Couples Pyrophosphatase Activity to DNA Replication |
title_short | Escherichia coli DnaE Polymerase Couples Pyrophosphatase Activity to DNA Replication |
title_sort | escherichia coli dnae polymerase couples pyrophosphatase activity to dna replication |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4822814/ https://www.ncbi.nlm.nih.gov/pubmed/27050298 http://dx.doi.org/10.1371/journal.pone.0152915 |
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