Journal
EMBO JOURNAL
Volume 32, Issue 9, Pages 1238-1249Publisher
NATURE PUBLISHING GROUP
DOI: 10.1038/emboj.2013.34
Keywords
bacterial chromosome segregation; diffusion ratchet; nucleoid; protein gradient; spatial organization
Categories
Funding
- Canadian Institutes of Health Research [37997]
- Program for Next Generation World-Leading Researchers [LS072]
- World Premier International Research Center Initiative (WPI) MEXT, Japan
- MEXT, Japan [23770204]
- Nancy Nossal Fellowship
- Grants-in-Aid for Scientific Research [23770204] Funding Source: KAKEN
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DNA segregation ensures the stable inheritance of genetic material prior to cell division. Many bacterial chromosomes and low-copy plasmids, such as the plasmids P1 and F, employ a three-component system to partition replicated genomes: a partition site on the DNA target, typically called parS, a partition site binding protein, typically called ParB, and a Walker-type ATPase, typically called ParA, which also binds non-specific DNA. In vivo, the ParA family of ATPases forms dynamic patterns over the nucleoid, but how ATP-driven patterning is involved in partition is unknown. We reconstituted and visualized ParA-mediated plasmid partition inside a DNA-carpeted flowcell, which acts as an artificial nucleoid. ParA and ParB transiently bridged plasmid to the DNA carpet. ParB-stimulated ATP hydrolysis by ParA resulted in ParA disassembly from the bridging complex and from the surrounding DNA carpet, which led to plasmid detachment. Our results support a diffusion-ratchet model, where ParB on the plasmid chases and redistributes the ParA gradient on the nucleoid, which in turn mobilizes the plasmid.
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