The WRN protein mutated in the premature aging disorder Werner syndrome is vital for
metabolism of perturbed replication forks. Replication Protein A (RPA) robustly enhances WRN
helicase activity in specific cases when tested in vitro. However, the significance of RPA-binding to
WRN in vivo has remained largely unexplored. We identified several conserved phosphorylation
sites in the acidic domain of WRN targeted by Casein Kinase 2 (CK2). These sites are crucial for
WRN-RPA interaction in vitro and in human cells. Using the CK2-unphosphorylatable WRN
mutant lacking the ability to bind RPA, we determined that the WRN-RPA complex plays a critical
role in fork recovery after replication stress whereas is not necessary for the processing of
replication forks or preventing DNA damage when forks stall or collapse. RPA-binding by WRN
and its helicase activity are crucial for countering the persistence of G4 structures after fork stalling.
Absence of WRN-RPA binding hampers fork recovery, causing single-strand DNA gaps, enlarged
by MRE11, and triggering MUS81-dependent double-strand breaks which requires efficient repair
by RAD51 to prevent excessive DNA damage.
[doi:10.25345/C55Q4RZ1Z]
[dataset license: CC0 1.0 Universal (CC0 1.0)]
Keywords: WERNER PHOSPHOSITES
Principal Investigators: (in alphabetical order) |
FEDERICA FRATINI, ISS, Italia |
Submitting User: | federica_fratini |
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