Defining the role of ATP hydrolysis in mitotic segregation of bacterial plasmids

PLoS Genetics
Yoan Ah-SengJean-Yves Bouet

Abstract

Hydrolysis of ATP by partition ATPases, although considered a key step in the segregation mechanism that assures stable inheritance of plasmids, is intrinsically very weak. The cognate centromere-binding protein (CBP), together with DNA, stimulates the ATPase to hydrolyse ATP and to undertake the relocation that incites plasmid movement, apparently confirming the need for hydrolysis in partition. However, ATP-binding alone changes ATPase conformation and properties, making it difficult to rigorously distinguish the substrate and cofactor roles of ATP in vivo. We had shown that mutation of arginines R36 and R42 in the F plasmid CBP, SopB, reduces stimulation of SopA-catalyzed ATP hydrolysis without changing SopA-SopB affinity, suggesting the role of hydrolysis could be analyzed using SopA with normal conformational responses to ATP. Here, we report that strongly reducing SopB-mediated stimulation of ATP hydrolysis results in only slight destabilization of mini-F, although the instability, as well as an increase in mini-F clustering, is proportional to the ATPase deficit. Unexpectedly, the reduced stimulation also increased the frequency of SopA relocation over the nucleoid. The increase was due to drastic shortening of the perio...Continue Reading

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Citations

Aug 12, 2014·Plasmid·Samuel Million-Weaver, Manel Camps
Dec 3, 2014·Current Opinion in Microbiology·Jean-Yves BouetFrançois Cornet
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Jan 12, 2021·The Journal of Membrane Biology·Dipika MishraRamanujam Srinivasan
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Jun 13, 2019·EcoSal Plus·Jean-Yves Bouet, Barbara E Funnell

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Methods Mentioned

BETA
fluorescence microscopy
PCR
electrophoresis

Software Mentioned

element
Image J
NIS
Metamorph

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