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Kinetic principles of ParA2-ATP cycling guide dynamic subcellular localizations in Vibrio cholerae

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posted on 2023-07-26, 16:13 authored by Satpal S Chodha, Adam C Brooks, Peter J Davis, Revathy Ramachandran, Dhruba K Chattoraj, Ling Chin Hwang
Abstract Dynamic protein gradients are exploited for the spatial organization and segregation of replicated chromosomes. However, mechanisms of protein gradient formation and how that spatially organizes chromosomes remain poorly understood. Here, we have determined the kinetic principles of subcellular localizations of ParA2 ATPase, an essential spatial regulator of chromosome 2 segregation in the multichromosome bacterium, Vibrio cholerae. We found that ParA2 gradients self-organize in V. cholerae cells into dynamic pole-to-pole oscillations. We examined the ParA2 ATPase cycle and ParA2 interactions with ParB2 and DNA. In vitro, ParA2-ATP dimers undergo a rate-limiting conformational switch, catalysed by DNA to achieve DNA-binding competence. This active ParA2 state loads onto DNA cooperatively as higher order oligomers. Our results indicate that the midcell localization of ParB2-parS2 complexes stimulate ATP hydrolysis and ParA2 release from the nucleoid, generating an asymmetric ParA2 gradient with maximal concentration toward the poles. This rapid dissociation coupled with slow nucleotide exchange and conformational switch provides for a temporal lag that allows the redistribution of ParA2 to the opposite pole for nucleoid reattachment. Based on our data, we propose a ‘Tug-of-war’ model that uses dynamic oscillations of ParA2 to spatially regulate symmetric segregation and positioning of bacterial chromosomes.

History

Refereed

  • Yes

Publication title

Nucleic Acids Research

ISSN

1362-4962

Publisher

Oxford University Press (OUP)

File version

  • Published version

Language

  • eng

Legacy posted date

2023-06-07

Legacy creation date

2023-06-07

Legacy Faculty/School/Department

Faculty of Health, Education, Medicine & Social Care

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