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The DnaK Chaperone Uses Different Mechanisms To Promote and Inhibit Replication of Vibrio cholerae Chromosome 2

Identifieur interne : 000B62 ( Main/Exploration ); précédent : 000B61; suivant : 000B63

The DnaK Chaperone Uses Different Mechanisms To Promote and Inhibit Replication of Vibrio cholerae Chromosome 2

Auteurs : Jyoti K. Jha [États-Unis] ; Mi Li [États-Unis] ; Rodolfo Ghirlando [États-Unis] ; Lisa M. Miller Jenkins [États-Unis] ; Alexander Wlodawer [États-Unis] ; Dhruba Chattoraj [États-Unis]

Source :

RBID : PMC:5395669

Descripteurs français

English descriptors

Abstract

ABSTRACT

Replication of Vibrio cholerae chromosome 2 (Chr2) depends on molecular chaperone DnaK to facilitate binding of the initiator (RctB) to the replication origin. The binding occurs at two kinds of site, 12-mers and 39-mers, which promote and inhibit replication, respectively. Here we show that DnaK employs different mechanisms to enhance the two kinds of binding. We found that mutations in rctB that reduce DnaK binding also reduce 12-mer binding and initiation. The initiation defect is suppressed by second-site mutations that increase 12-mer binding only marginally. Instead, they reduce replication inhibitory mechanisms: RctB dimerization and 39-mer binding. One suppressing change was in a dimerization domain which is folded similarly to the initiator of an iteron plasmid—the presumed progenitor of Chr2. In plasmids, DnaK promotes initiation by reducing dimerization. A different mutation was in the 39-mer binding domain of RctB and inactivated it, indicating an alternative suppression mechanism. Paradoxically, although DnaK increases 39-mer binding, the increase was also achieved by inactivating the DnaK binding site of RctB. This result suggests that the site inhibits the 39-mer binding domain (via autoinhibition) when prevented from binding DnaK. Taken together, our results reveal an important feature of the transition from plasmid to chromosome: the Chr2 initiator retains the plasmid-like dimerization domain and its control by chaperones but uses the chaperones in an unprecedented way to control the inhibitory 39-mer binding.


Url:
DOI: 10.1128/mBio.00427-17
PubMed: 28420739
PubMed Central: 5395669


Affiliations:


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Le document en format XML

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<term>Origine de réplication</term>
<term>Protéines bactériennes (métabolisme)</term>
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<p>Replication of
<italic>Vibrio cholerae</italic>
chromosome 2 (Chr2) depends on molecular chaperone DnaK to facilitate binding of the initiator (RctB) to the replication origin. The binding occurs at two kinds of site, 12-mers and 39-mers, which promote and inhibit replication, respectively. Here we show that DnaK employs different mechanisms to enhance the two kinds of binding. We found that mutations in
<italic>rctB</italic>
that reduce DnaK binding also reduce 12-mer binding and initiation. The initiation defect is suppressed by second-site mutations that increase 12-mer binding only marginally. Instead, they reduce replication inhibitory mechanisms: RctB dimerization and 39-mer binding. One suppressing change was in a dimerization domain which is folded similarly to the initiator of an iteron plasmid—the presumed progenitor of Chr2. In plasmids, DnaK promotes initiation by reducing dimerization. A different mutation was in the 39-mer binding domain of RctB and inactivated it, indicating an alternative suppression mechanism. Paradoxically, although DnaK increases 39-mer binding, the increase was also achieved by inactivating the DnaK binding site of RctB. This result suggests that the site inhibits the 39-mer binding domain (via autoinhibition) when prevented from binding DnaK. Taken together, our results reveal an important feature of the transition from plasmid to chromosome: the Chr2 initiator retains the plasmid-like dimerization domain and its control by chaperones but uses the chaperones in an unprecedented way to control the inhibitory 39-mer binding.</p>
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<name sortKey="Grosse Kunstleve, Rw" uniqKey="Grosse Kunstleve R">RW Grosse-Kunstleve</name>
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<name sortKey="Hung, Lw" uniqKey="Hung L">LW Hung</name>
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<name sortKey="Ioerger, Tr" uniqKey="Ioerger T">TR Ioerger</name>
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<author>
<name sortKey="Mccoy, Aj" uniqKey="Mccoy A">AJ McCoy</name>
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<author>
<name sortKey="Moriarty, Nw" uniqKey="Moriarty N">NW Moriarty</name>
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<author>
<name sortKey="Read, Rj" uniqKey="Read R">RJ Read</name>
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<name sortKey="Sacchettini, Jc" uniqKey="Sacchettini J">JC Sacchettini</name>
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<author>
<name sortKey="Sauter, Nk" uniqKey="Sauter N">NK Sauter</name>
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<name sortKey="Terwilliger, Tc" uniqKey="Terwilliger T">TC Terwilliger</name>
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<author>
<name sortKey="Venkova Canova, T" uniqKey="Venkova Canova T">T Venkova-Canova</name>
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<author>
<name sortKey="Srivastava, P" uniqKey="Srivastava P">P Srivastava</name>
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<author>
<name sortKey="Chattoraj, Dk" uniqKey="Chattoraj D">DK Chattoraj</name>
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<li>Maryland</li>
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<name sortKey="Jha, Jyoti K" sort="Jha, Jyoti K" uniqKey="Jha J" first="Jyoti K." last="Jha">Jyoti K. Jha</name>
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<name sortKey="Chattoraj, Dhruba" sort="Chattoraj, Dhruba" uniqKey="Chattoraj D" first="Dhruba" last="Chattoraj">Dhruba Chattoraj</name>
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<name sortKey="Miller Jenkins, Lisa M" sort="Miller Jenkins, Lisa M" uniqKey="Miller Jenkins L" first="Lisa M." last="Miller Jenkins">Lisa M. Miller Jenkins</name>
<name sortKey="Wlodawer, Alexander" sort="Wlodawer, Alexander" uniqKey="Wlodawer A" first="Alexander" last="Wlodawer">Alexander Wlodawer</name>
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