TY - JOUR
T1 - A local ATR-dependent checkpoint pathway is activated by a site-specific replication fork block in human cells
AU - Ahmed-Seghir, Sana
AU - Jalan, Manisha
AU - Grimsley, Helen E.
AU - Sharma, Aman
AU - Twayana, Shyam
AU - Kosiyatrakul, Settapong T.
AU - Thompson, Christopher
AU - Schildkraut, Carl L.
AU - Powell, Simon N.
N1 - Publisher Copyright:
© Ahmed-Seghir, Jalan et al.
PY - 2023
Y1 - 2023
N2 - When replication forks encounter DNA lesions that cause polymerase stalling, a checkpoint pathway is activated. The ATR-dependent intra-S checkpoint pathway mediates detection and processing of sites of replication fork stalling to maintain genomic integrity. Several factors involved in the global checkpoint pathway have been identified, but the response to a single replication fork barrier (RFB) is poorly understood. We utilized the Escherichia coli-based Tus-Ter system in human MCF7 cells and showed that the Tus protein binding to TerB sequences creates an efficient site-specific RFB. The single fork RFB was sufficient to activate a local, but not global, ATR-dependent checkpoint response that leads to phosphorylation and accumulation of DNA damage sensor protein γH2AX, confined locally to within a kilobase of the site of stalling. These data support a model of local management of fork stalling, which allows global replication at sites other than the RFB to continue to progress without delay.
AB - When replication forks encounter DNA lesions that cause polymerase stalling, a checkpoint pathway is activated. The ATR-dependent intra-S checkpoint pathway mediates detection and processing of sites of replication fork stalling to maintain genomic integrity. Several factors involved in the global checkpoint pathway have been identified, but the response to a single replication fork barrier (RFB) is poorly understood. We utilized the Escherichia coli-based Tus-Ter system in human MCF7 cells and showed that the Tus protein binding to TerB sequences creates an efficient site-specific RFB. The single fork RFB was sufficient to activate a local, but not global, ATR-dependent checkpoint response that leads to phosphorylation and accumulation of DNA damage sensor protein γH2AX, confined locally to within a kilobase of the site of stalling. These data support a model of local management of fork stalling, which allows global replication at sites other than the RFB to continue to progress without delay.
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U2 - 10.7554/eLife.87357
DO - 10.7554/eLife.87357
M3 - Article
C2 - 37647215
AN - SCOPUS:85169230001
SN - 2050-084X
VL - 12
JO - eLife
JF - eLife
M1 - RP87357
ER -