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2015 ; 59
(6
): 998-1010
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English Wikipedia
The Replication Checkpoint Prevents Two Types of Fork Collapse without Regulating
Replisome Stability
#MMPMID26365379
Dungrawala H
; Rose KL
; Bhat KP
; Mohni KN
; Glick GG
; Couch FB
; Cortez D
Mol Cell
2015[Sep]; 59
(6
): 998-1010
PMID26365379
show ga
The ATR replication checkpoint ensures that stalled forks remain stable when
replisome movement is impeded. Using an improved iPOND protocol combined with
SILAC mass spectrometry, we characterized human replisome dynamics in response to
fork stalling. Our data provide a quantitative picture of the replisome and
replication stress response proteomes in 32 experimental conditions. Importantly,
rather than stabilize the replisome, the checkpoint prevents two distinct types
of fork collapse. Unsupervised hierarchical clustering of protein abundance on
nascent DNA is sufficient to identify protein complexes and place newly
identified replisome-associated proteins into functional pathways. As an example,
we demonstrate that ZNF644 complexes with the G9a/GLP methyltransferase at
replication forks and is needed to prevent replication-associated DNA damage. Our
data reveal how the replication checkpoint preserves genome integrity, provide
insights into the mechanism of action of ATR inhibitors, and will be a useful
resource for replication, DNA repair, and chromatin investigators.