Dynamic Distribution of SeqA Protein across the Chromosome of <named-content content-type="genus-species">Escherichia coli</named-content> K-12

ABSTRACT The bacterial SeqA protein binds to hemi-methylated GATC sequences that arise in newly synthesized DNA upon passage of the replication machinery. In Escherichia coli K-12, the single replication origin oriC is a well-characterized target for SeqA, which binds to multiple hemi-methylated GAT...

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Autores principales:	María Antonia Sánchez-Romero, Stephen J. W. Busby, Nigel P. Dyer, Sascha Ott, Andrew D. Millard, David C. Grainger
Formato:	article
Lenguaje:	EN
Publicado:	American Society for Microbiology 2010
Materias:	Microbiology QR1-502
Acceso en línea:	https://doaj.org/article/059bbc1f4514407cbb3e0a2c163f2460
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spelling	oai:doaj.org-article:059bbc1f4514407cbb3e0a2c163f24602021-11-15T15:38:14ZDynamic Distribution of SeqA Protein across the Chromosome of <named-content content-type="genus-species">Escherichia coli</named-content> K-1210.1128/mBio.00012-102150-7511https://doaj.org/article/059bbc1f4514407cbb3e0a2c163f24602010-05-01T00:00:00Zhttps://journals.asm.org/doi/10.1128/mBio.00012-10https://doaj.org/toc/2150-7511ABSTRACT The bacterial SeqA protein binds to hemi-methylated GATC sequences that arise in newly synthesized DNA upon passage of the replication machinery. In Escherichia coli K-12, the single replication origin oriC is a well-characterized target for SeqA, which binds to multiple hemi-methylated GATC sequences immediately after replication has initiated. This sequesters oriC, thereby preventing reinitiation of replication. However, the genome-wide DNA binding properties of SeqA are unknown, and hence, here, we describe a study of the binding of SeqA across the entire Escherichia coli K-12 chromosome, using chromatin immunoprecipitation in combination with DNA microarrays. Our data show that SeqA binding correlates with the frequency and spacing of GATC sequences across the entire genome. Less SeqA is found in highly transcribed regions, as well as in the ter macrodomain. Using synchronized cultures, we show that SeqA distribution differs with the cell cycle. SeqA remains bound to some targets after replication has ceased, and these targets locate to genes encoding factors involved in nucleotide metabolism, chromosome replication, and methyl transfer. IMPORTANCE DNA replication in bacteria is a highly regulated process. In many bacteria, a protein called SeqA plays a key role by binding to newly replicated DNA. Thus, at the origin of DNA replication, SeqA binding blocks premature reinitiation of replication rounds. Although most investigators have focused on the role of SeqA at replication origins, it has long been suspected that SeqA has a more pervasive role. In this study, we describe how we have been able to identify scores of targets, across the entire Escherichia coli chromosome, to which SeqA binds. Using synchronously growing cells, we show that the distribution of SeqA between these targets alters as replication of the chromosome progresses. This suggests that sequential changes in SeqA distribution orchestrate a program of gene expression that ensures coordinated DNA replication and cell division.María Antonia Sánchez-RomeroStephen J. W. BusbyNigel P. DyerSascha OttAndrew D. MillardDavid C. GraingerAmerican Society for MicrobiologyarticleMicrobiologyQR1-502ENmBio, Vol 1, Iss 1 (2010)
institution	DOAJ
collection	DOAJ
language	EN
topic	Microbiology QR1-502
spellingShingle	Microbiology QR1-502 María Antonia Sánchez-Romero Stephen J. W. Busby Nigel P. Dyer Sascha Ott Andrew D. Millard David C. Grainger Dynamic Distribution of SeqA Protein across the Chromosome of <named-content content-type="genus-species">Escherichia coli</named-content> K-12
description	ABSTRACT The bacterial SeqA protein binds to hemi-methylated GATC sequences that arise in newly synthesized DNA upon passage of the replication machinery. In Escherichia coli K-12, the single replication origin oriC is a well-characterized target for SeqA, which binds to multiple hemi-methylated GATC sequences immediately after replication has initiated. This sequesters oriC, thereby preventing reinitiation of replication. However, the genome-wide DNA binding properties of SeqA are unknown, and hence, here, we describe a study of the binding of SeqA across the entire Escherichia coli K-12 chromosome, using chromatin immunoprecipitation in combination with DNA microarrays. Our data show that SeqA binding correlates with the frequency and spacing of GATC sequences across the entire genome. Less SeqA is found in highly transcribed regions, as well as in the ter macrodomain. Using synchronized cultures, we show that SeqA distribution differs with the cell cycle. SeqA remains bound to some targets after replication has ceased, and these targets locate to genes encoding factors involved in nucleotide metabolism, chromosome replication, and methyl transfer. IMPORTANCE DNA replication in bacteria is a highly regulated process. In many bacteria, a protein called SeqA plays a key role by binding to newly replicated DNA. Thus, at the origin of DNA replication, SeqA binding blocks premature reinitiation of replication rounds. Although most investigators have focused on the role of SeqA at replication origins, it has long been suspected that SeqA has a more pervasive role. In this study, we describe how we have been able to identify scores of targets, across the entire Escherichia coli chromosome, to which SeqA binds. Using synchronously growing cells, we show that the distribution of SeqA between these targets alters as replication of the chromosome progresses. This suggests that sequential changes in SeqA distribution orchestrate a program of gene expression that ensures coordinated DNA replication and cell division.
format	article
author	María Antonia Sánchez-Romero Stephen J. W. Busby Nigel P. Dyer Sascha Ott Andrew D. Millard David C. Grainger
author_facet	María Antonia Sánchez-Romero Stephen J. W. Busby Nigel P. Dyer Sascha Ott Andrew D. Millard David C. Grainger
author_sort	María Antonia Sánchez-Romero
title	Dynamic Distribution of SeqA Protein across the Chromosome of <named-content content-type="genus-species">Escherichia coli</named-content> K-12
title_short	Dynamic Distribution of SeqA Protein across the Chromosome of <named-content content-type="genus-species">Escherichia coli</named-content> K-12
title_full	Dynamic Distribution of SeqA Protein across the Chromosome of <named-content content-type="genus-species">Escherichia coli</named-content> K-12
title_fullStr	Dynamic Distribution of SeqA Protein across the Chromosome of <named-content content-type="genus-species">Escherichia coli</named-content> K-12
title_full_unstemmed	Dynamic Distribution of SeqA Protein across the Chromosome of <named-content content-type="genus-species">Escherichia coli</named-content> K-12
title_sort	dynamic distribution of seqa protein across the chromosome of <named-content content-type="genus-species">escherichia coli</named-content> k-12
publisher	American Society for Microbiology
publishDate	2010
url	https://doaj.org/article/059bbc1f4514407cbb3e0a2c163f2460
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Dynamic Distribution of SeqA Protein across the Chromosome of <named-content content-type="genus-species">Escherichia coli</named-content> K-12

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