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C-jun n-terminal kinase-mediated rad18 phosphorylation facilitates pol  recruitment to stalled replication forks

Barkley, L. R.
; 
Palle, K.
; 
Durando, M.
; 
Day, T. A.
; 
Gurkar, A.
; 
Kakusho, N.
; 
Li, J.
; 
Masai, H.
; 
Vaziri, C.
Repository DOI
Publication Date
2012-03-28
Keywords
DNA-polymerase-eta, s-phase checkpoint, strand break repair, xeroderma-pigmentosum, signal-transduction, protein-a, pcna monoubiquitination, translesion synthesis, ultraviolet-light, damage bypass
Type
Article
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Citation
Barkley, L. R. Palle, K.; Durando, M.; Day, T. A.; Gurkar, A.; Kakusho, N.; Li, J.; Masai, H.; Vaziri, C. (2012). C-jun n-terminal kinase-mediated rad18 phosphorylation facilitates pol  recruitment to stalled replication forks. Molecular Biology of the Cell 23 (10), 1943-1954
Abstract
The E3 ubiquitin ligase Rad18 chaperones DNA polymerase eta (Pol eta) to sites of UV-induced DNA damage and monoubiquitinates proliferating cell nuclear antigen (PCNA), facilitating engagement of Pol eta with stalled replication forks and promoting translesion synthesis (TLS). It is unclear how Rad18 activities are coordinated with other elements of the DNA damage response. We show here that Ser-409 residing in the Pol eta-binding motif of Rad18 is phosphorylated in a checkpoint kinase 1-dependent manner in genotoxin-treated cells. Recombinant Rad18 was phosphorylated specifically at S409 by c-Jun N-terminal kinase (JNK) in vitro. In UV-treated cells, Rad18 S409 phosphorylation was inhibited by a pharmacological JNK inhibitor. Conversely, ectopic expression of JNK and its upstream kinase mitogen-activated protein kinase kinase 4 led to DNA damage-independent Rad18 S409 phosphorylation. These results identify Rad18 as a novel JNK substrate. A Rad18 mutant harboring a Ser -> Ala substitution at S409 was compromised for Pol. association and did not redistribute Pol eta to nuclear foci or promote Pol eta-PCNA interaction efficiently relative to wild-type Rad18. Rad18 S409A also failed to fully complement the UV sensitivity of Rad18-depleted cells. Taken together, these results show that Rad18 phosphorylation by JNK represents a novel mechanism for promoting TLS and DNA damage tolerance.
Funder
Publisher
American Society for Cell Biology (ASCB)
Publisher DOI
10.1091/mbc.e11-10-0829
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Attribution-NonCommercial-NoDerivs 3.0 Ireland
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Externally hosted open access publications with University of Galway authors (2)