Gyrase and Topo IV modulate chromosome domain size in vivo.

Abstract:

:In bacteria, DNA supercoil movement is restricted to subchromosomal regions or 'domains.' To elucidate the nature of domain boundaries, we analysed reaction kinetics for gammadelta site-specific resolution in six chromosomal intervals ranging in size from 14 to 90 kb. In stationary cultures of Salmonella typhimurium, resolution kinetics were rapid for both short and long intervals, suggesting that random stationary barriers occur with a 30% probability at approximately 80 kb intervals along DNA. To test the biochemical nature of domain barriers, a genetic screen was used to look for mutants with small domains. Rare temperature-sensitive alleles of DNA gyrase and Topo IV (the two essential type II topoisomerases) had more supercoil barriers than wild-type strains in all growth states. The most severe gyrase mutants were found to have twice as many barriers in growing cells as wild type throughout a 90 kb interval of the chromosome. We propose that knots and tangles in duplex DNA restrain supercoil diffusion in living bacteria.

journal_name

Mol Microbiol

journal_title

Molecular microbiology

authors

Staczek P,Higgins NP

doi

10.1046/j.1365-2958.1998.01025.x

subject

Has Abstract

pub_date

1998-09-01 00:00:00

pages

1435-48

issue

6

eissn

0950-382X

issn

1365-2958

journal_volume

29

pub_type

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