Two mechanisms of chromosome fragility at replication-termination sites in bacteria

Chromosomal fragile sites are implicated in promoting genome instability, which drives cancers and neurological diseases. Yet, the causes and mechanisms of chromosome fragility remain speculative. Here, we identify three spontaneous fragile sites in the Escherichia coli genome and define their DNA damage and repair intermediates at high resolution. We find that all three sites, all in the region of replication termination, display recurrent four-way DNA or Holliday junctions (HJs) and recurrent DNA breaks. Homology-directed double-strand break repair generates the recurrent HJs at all of these sites; however, distinct mechanisms of DNA breakage are implicated: replication fork collapse at natural replication barriers and, unexpectedly, frequent shearing of unsegregated sister chromosomes at cell division. We propose that mechanisms such as both of these may occur ubiquitously, including in humans, and may constitute some of the earliest events that underlie somatic cell mosaicism, cancers, and other diseases of genome instability.

Automated summary

Chromosomal fragile sites are implicated in promoting genome instability, and this study identifies three spontaneous fragile sites in the Escherichia coli genome, revealing their DNA damage and repair mechanisms. The findings suggest that mechanisms such as replication fork collapse and shearing of unsegregated sister chromosomes may occur universally, including in humans, and play a role in diseases of genome instability such as somatic cell mosaicism and cancers.