Association of the chromosome replication initiator DnaA with the Escherichia coli inner membrane in vivo: quantity and mode of binding.

DnaA initiates chromosome replication in most known bacteria and its activity is controlled so that this event occurs only once every cell division cycle. ATP in the active ATP-DnaA is hydrolyzed after initiation and the resulting ADP is replaced with ATP on the verge of the next initiation. Two put...

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Autores principales:	Tomer Regev, Nadav Myers, Raz Zarivach, Itzhak Fishov
Formato:	article
Lenguaje:	EN
Publicado:	Public Library of Science (PLoS) 2012
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/3b355e88cfbe43e0b448ca6082ca80c9
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spelling	oai:doaj.org-article:3b355e88cfbe43e0b448ca6082ca80c92021-11-18T07:19:38ZAssociation of the chromosome replication initiator DnaA with the Escherichia coli inner membrane in vivo: quantity and mode of binding.1932-620310.1371/journal.pone.0036441https://doaj.org/article/3b355e88cfbe43e0b448ca6082ca80c92012-01-01T00:00:00Zhttps://www.ncbi.nlm.nih.gov/pmc/articles/pmid/22574163/pdf/?tool=EBIhttps://doaj.org/toc/1932-6203DnaA initiates chromosome replication in most known bacteria and its activity is controlled so that this event occurs only once every cell division cycle. ATP in the active ATP-DnaA is hydrolyzed after initiation and the resulting ADP is replaced with ATP on the verge of the next initiation. Two putative recycling mechanisms depend on the binding of DnaA either to the membrane or to specific chromosomal sites, promoting nucleotide dissociation. While there is no doubt that DnaA interacts with artificial membranes in vitro, it is still controversial as to whether it binds the cytoplasmic membrane in vivo. In this work we looked for DnaA-membrane interaction in E. coli cells by employing cell fractionation with both native and fluorescent DnaA hybrids. We show that about 10% of cellular DnaA is reproducibly membrane-associated. This small fraction might be physiologically significant and represent the free DnaA available for initiation, rather than the vast majority bound to the datA reservoir. Using the combination of mCherry with a variety of DnaA fragments, we demonstrate that the membrane binding function is delocalized on the surface of the protein's domain III, rather than confined to a particular sequence. We propose a new binding-bending mechanism to explain the membrane-induced nucleotide release from DnaA. This mechanism would be fundamental to the initiation of replication.Tomer RegevNadav MyersRaz ZarivachItzhak FishovPublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 7, Iss 5, p e36441 (2012)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Tomer Regev Nadav Myers Raz Zarivach Itzhak Fishov Association of the chromosome replication initiator DnaA with the Escherichia coli inner membrane in vivo: quantity and mode of binding.
description	DnaA initiates chromosome replication in most known bacteria and its activity is controlled so that this event occurs only once every cell division cycle. ATP in the active ATP-DnaA is hydrolyzed after initiation and the resulting ADP is replaced with ATP on the verge of the next initiation. Two putative recycling mechanisms depend on the binding of DnaA either to the membrane or to specific chromosomal sites, promoting nucleotide dissociation. While there is no doubt that DnaA interacts with artificial membranes in vitro, it is still controversial as to whether it binds the cytoplasmic membrane in vivo. In this work we looked for DnaA-membrane interaction in E. coli cells by employing cell fractionation with both native and fluorescent DnaA hybrids. We show that about 10% of cellular DnaA is reproducibly membrane-associated. This small fraction might be physiologically significant and represent the free DnaA available for initiation, rather than the vast majority bound to the datA reservoir. Using the combination of mCherry with a variety of DnaA fragments, we demonstrate that the membrane binding function is delocalized on the surface of the protein's domain III, rather than confined to a particular sequence. We propose a new binding-bending mechanism to explain the membrane-induced nucleotide release from DnaA. This mechanism would be fundamental to the initiation of replication.
format	article
author	Tomer Regev Nadav Myers Raz Zarivach Itzhak Fishov
author_facet	Tomer Regev Nadav Myers Raz Zarivach Itzhak Fishov
author_sort	Tomer Regev
title	Association of the chromosome replication initiator DnaA with the Escherichia coli inner membrane in vivo: quantity and mode of binding.
title_short	Association of the chromosome replication initiator DnaA with the Escherichia coli inner membrane in vivo: quantity and mode of binding.
title_full	Association of the chromosome replication initiator DnaA with the Escherichia coli inner membrane in vivo: quantity and mode of binding.
title_fullStr	Association of the chromosome replication initiator DnaA with the Escherichia coli inner membrane in vivo: quantity and mode of binding.
title_full_unstemmed	Association of the chromosome replication initiator DnaA with the Escherichia coli inner membrane in vivo: quantity and mode of binding.
title_sort	association of the chromosome replication initiator dnaa with the escherichia coli inner membrane in vivo: quantity and mode of binding.
publisher	Public Library of Science (PLoS)
publishDate	2012
url	https://doaj.org/article/3b355e88cfbe43e0b448ca6082ca80c9
work_keys_str_mv	AT tomerregev associationofthechromosomereplicationinitiatordnaawiththeescherichiacoliinnermembraneinvivoquantityandmodeofbinding AT nadavmyers associationofthechromosomereplicationinitiatordnaawiththeescherichiacoliinnermembraneinvivoquantityandmodeofbinding AT razzarivach associationofthechromosomereplicationinitiatordnaawiththeescherichiacoliinnermembraneinvivoquantityandmodeofbinding AT itzhakfishov associationofthechromosomereplicationinitiatordnaawiththeescherichiacoliinnermembraneinvivoquantityandmodeofbinding
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Association of the chromosome replication initiator DnaA with the Escherichia coli inner membrane in vivo: quantity and mode of binding.

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