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Cooperative DnaA Binding to the Negatively Supercoiled datA Locus Stimulates DnaA-ATP Hydrolysis.

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Title: Cooperative DnaA Binding to the Negatively Supercoiled datA Locus Stimulates DnaA-ATP Hydrolysis.
Authors: Kazutoshi Kasho1, Hiroyuki Tanaka1, Ryuji Sakai1, Tsutomu Katayama1 katayama@phar.kyushu-u.ac.jp
Source: Journal of Biological Chemistry. 1/27/2017, Vol. 292 Issue 4, p1251-1266. 16p.
Subjects: DNA analysis, Integration host factor, Bacterial proteins, Novobiocin, Fluoroquinolones
Abstract: Timely initiation of replication in Escherichia coli requires functional regulation of the replication initiator, ATP-DnaA. The cellular level of ATP-DnaA increases just before initiation, after which its level decreases through hydrolysis of DnaA-bound ATP, yielding initiation-inactive ADP-DnaA. Previously, we reported a novel DnaA-ATP hydrolysis system involving the chromosomal locus datA and named it datA-dependent DnaA-ATP hydrolysis (DDAH). The datA locus contains a binding site for a nucleoid-associating factor integration host factor (IHF) and a cluster of three known DnaA-binding sites, which are important for DDAH. However, the mechanisms underlying the formation and regulation of the datA-IHF·DnaA complex remain unclear. We now demonstrate that a novel DnaA box within datA is essential for ATP-DnaA complex formation and DnaA-ATP hydrolysis. Specific DnaA residues, which are important for interaction with bound ATP and for head-to-tail inter-DnaA interaction, were also required for ATP-DnaA-specific oligomer formation on datA. Furthermore, we show that negative DNA supercoiling of datA stabilizes ATP-DnaA oligomers, and stimulates datA-IHF interaction and DnaA-ATP hydrolysis. Relaxation of DNA supercoiling by the addition of novobiocin, a DNA gyrase inhibitor, inhibits datA function in cells. On the basis of these results, we propose a mechanistic model of datA-IHF·DnaA complex formation and DNA supercoiling-dependent regulation for DDAH. [ABSTRACT FROM AUTHOR]
Copyright of Journal of Biological Chemistry is the property of Elsevier B.V. and its content may not be copied or emailed to multiple sites without the copyright holder's express written permission. Additionally, content may not be used with any artificial intelligence tools or machine learning technologies. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)
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DbLabel: Engineering Source
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  Label: Title
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  Data: Cooperative DnaA Binding to the Negatively Supercoiled datA Locus Stimulates DnaA-ATP Hydrolysis.
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  Data: <searchLink fieldCode="AR" term="%22Kazutoshi+Kasho%22">Kazutoshi Kasho</searchLink><relatesTo>1</relatesTo><br /><searchLink fieldCode="AR" term="%22Hiroyuki+Tanaka%22">Hiroyuki Tanaka</searchLink><relatesTo>1</relatesTo><br /><searchLink fieldCode="AR" term="%22Ryuji+Sakai%22">Ryuji Sakai</searchLink><relatesTo>1</relatesTo><br /><searchLink fieldCode="AR" term="%22Tsutomu+Katayama%22">Tsutomu Katayama</searchLink><relatesTo>1</relatesTo><i> katayama@phar.kyushu-u.ac.jp</i>
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  Data: <searchLink fieldCode="JN" term="%22Journal+of+Biological+Chemistry%22">Journal of Biological Chemistry</searchLink>. 1/27/2017, Vol. 292 Issue 4, p1251-1266. 16p.
– Name: Subject
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  Data: <searchLink fieldCode="DE" term="%22DNA+analysis%22">DNA analysis</searchLink><br /><searchLink fieldCode="DE" term="%22Integration+host+factor%22">Integration host factor</searchLink><br /><searchLink fieldCode="DE" term="%22Bacterial+proteins%22">Bacterial proteins</searchLink><br /><searchLink fieldCode="DE" term="%22Novobiocin%22">Novobiocin</searchLink><br /><searchLink fieldCode="DE" term="%22Fluoroquinolones%22">Fluoroquinolones</searchLink>
– Name: Abstract
  Label: Abstract
  Group: Ab
  Data: Timely initiation of replication in Escherichia coli requires functional regulation of the replication initiator, ATP-DnaA. The cellular level of ATP-DnaA increases just before initiation, after which its level decreases through hydrolysis of DnaA-bound ATP, yielding initiation-inactive ADP-DnaA. Previously, we reported a novel DnaA-ATP hydrolysis system involving the chromosomal locus datA and named it datA-dependent DnaA-ATP hydrolysis (DDAH). The datA locus contains a binding site for a nucleoid-associating factor integration host factor (IHF) and a cluster of three known DnaA-binding sites, which are important for DDAH. However, the mechanisms underlying the formation and regulation of the datA-IHF·DnaA complex remain unclear. We now demonstrate that a novel DnaA box within datA is essential for ATP-DnaA complex formation and DnaA-ATP hydrolysis. Specific DnaA residues, which are important for interaction with bound ATP and for head-to-tail inter-DnaA interaction, were also required for ATP-DnaA-specific oligomer formation on datA. Furthermore, we show that negative DNA supercoiling of datA stabilizes ATP-DnaA oligomers, and stimulates datA-IHF interaction and DnaA-ATP hydrolysis. Relaxation of DNA supercoiling by the addition of novobiocin, a DNA gyrase inhibitor, inhibits datA function in cells. On the basis of these results, we propose a mechanistic model of datA-IHF·DnaA complex formation and DNA supercoiling-dependent regulation for DDAH. [ABSTRACT FROM AUTHOR]
– Name: AbstractSuppliedCopyright
  Label:
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  Data: <i>Copyright of Journal of Biological Chemistry is the property of Elsevier B.V. and its content may not be copied or emailed to multiple sites without the copyright holder's express written permission. Additionally, content may not be used with any artificial intelligence tools or machine learning technologies. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract.</i> (Copyright applies to all Abstracts.)
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      – Type: doi
        Value: 10.1074/jbc.M116.762815
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      – Code: eng
        Text: English
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        PageCount: 16
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      – SubjectFull: DNA analysis
        Type: general
      – SubjectFull: Integration host factor
        Type: general
      – SubjectFull: Bacterial proteins
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      – SubjectFull: Novobiocin
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      – SubjectFull: Fluoroquinolones
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      – TitleFull: Cooperative DnaA Binding to the Negatively Supercoiled datA Locus Stimulates DnaA-ATP Hydrolysis.
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            NameFull: Kazutoshi Kasho
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            NameFull: Hiroyuki Tanaka
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            NameFull: Ryuji Sakai
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            NameFull: Tsutomu Katayama
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            – D: 27
              M: 01
              Text: 1/27/2017
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              Y: 2017
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