MRX protects fork integrity at protein-DNA barriers, and its absence causes checkpoint activation dependent on chromatin context.

Abstract:

:To address how eukaryotic replication forks respond to fork stalling caused by strong non-covalent protein-DNA barriers, we engineered the controllable Fob-block system in Saccharomyces cerevisiae. This system allows us to strongly induce and control replication fork barriers (RFB) at their natural location within the rDNA. We discover a pivotal role for the MRX (Mre11, Rad50, Xrs2) complex for fork integrity at RFBs, which differs from its acknowledged function in double-strand break processing. Consequently, in the absence of the MRX complex, single-stranded DNA (ssDNA) accumulates at the rDNA. Based on this, we propose a model where the MRX complex specifically protects stalled forks at protein-DNA barriers, and its absence leads to processing resulting in ssDNA. To our surprise, this ssDNA does not trigger a checkpoint response. Intriguingly, however, placing RFBs ectopically on chromosome VI provokes a strong Rad53 checkpoint activation in the absence of Mre11. We demonstrate that proper checkpoint signalling within the rDNA is restored on deletion of SIR2. This suggests the surprising and novel concept that chromatin is an important player in checkpoint signalling.

journal_name

Nucleic Acids Res

journal_title

Nucleic acids research

authors

Bentsen IB,Nielsen I,Lisby M,Nielsen HB,Gupta SS,Mundbjerg K,Andersen AH,Bjergbaek L

doi

10.1093/nar/gkt051

subject

Has Abstract

pub_date

2013-03-01 00:00:00

pages

3173-89

issue

5

eissn

0305-1048

issn

1362-4962

pii

gkt051

journal_volume

41

pub_type

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