High-resolution mapping of mitotic DNA synthesis regions and common fragile sites in the human genome through direct sequencing.

Abstract:

:DNA replication stress, a feature of human cancers, often leads to instability at specific genomic loci, such as the common fragile sites (CFSs). Cells experiencing DNA replication stress may also exhibit mitotic DNA synthesis (MiDAS). To understand the physiological function of MiDAS and its relationship to CFSs, we mapped, at high resolution, the genomic sites of MiDAS in cells treated with the DNA polymerase inhibitor aphidicolin. Sites of MiDAS were evident as well-defined peaks that were largely conserved between cell lines and encompassed all known CFSs. The MiDAS peaks mapped within large, transcribed, origin-poor genomic regions. In cells that had been treated with aphidicolin, these regions remained unreplicated even in late S phase; MiDAS then served to complete their replication after the cells entered mitosis. Interestingly, leading and lagging strand synthesis were uncoupled in MiDAS, consistent with MiDAS being a form of break-induced replication, a repair mechanism for collapsed DNA replication forks. Our results provide a better understanding of the mechanisms leading to genomic instability at CFSs and in cancer cells.

journal_name

Cell Res

journal_title

Cell research

authors

Macheret M,Bhowmick R,Sobkowiak K,Padayachy L,Mailler J,Hickson ID,Halazonetis TD

doi

10.1038/s41422-020-0358-x

subject

Has Abstract

pub_date

2020-11-01 00:00:00

pages

997-1008

issue

11

eissn

1001-0602

issn

1748-7838

pii

10.1038/s41422-020-0358-x

journal_volume

30

pub_type

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