Other Genome binding/occupancy profiling by high throughput sequencing
Summary
SMC condensin complexes play a central role in compacting and resolving replicated chromosomes in virtually all organisms yet how they accomplish this remains elusive. In Bacillus subtilis, condensin is loaded at centromeric parS sites, where it encircles DNA and individualizes newly replicated origins. Using chromosome conformation capture and cytological assays, we show that condensin recruitment to origin-proximal parS sites is required for the juxtaposition of the two chromosome arms. Recruitment to ectopic parS sites promotes alignment of large tracks of DNA flanking these sites. Importantly, insertion of parS sites on opposing arms indicates that these ?zip-up? interactions only occur between adjacent DNA segments. Collectively, our data suggest that condensin resolves replicated origins by promoting the juxtaposition of DNA flanking parS sites drawing sister origins in on themselves and away from each other. These results are consistent with a model in which condensin encircles the DNA flanking its loading site and then slides down, tethering the two arms together. Lengthwise condensation via loop-extrusion could provide a generalizable mechanism by which condensin acts dynamically to individualize origins in B. subtilis and, when loaded along eukaryotic chromosomes, to resolve them during mitosis.
Overall design
Hi-C experiments were performed on wild type and mutant cells of Bacillus subtilis PY79 growing in rich or poor media. ChIP-seq experiments were performed on two strains of Bacillus subtilis PY79 growing in the same condition as in Hi-C experiments.
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