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c-Jun N-terminal kinase–mediated Rad18 phosphorylation facilitates Polη recruitment to stalled replication forks

The E3 ubiquitin ligase Rad18 chaperones DNA polymerase η (Polη) to sites of UV-induced DNA damage and monoubiquitinates proliferating cell nuclear antigen (PCNA), facilitating engagement of Polη with stalled replication forks and promoting translesion synthesis (TLS). It is unclear how Rad18 activi...

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Autores principales: Barkley, Laura R., Palle, Komaraiah, Durando, Michael, Day, Tovah A., Gurkar, Aditi, Kakusho, Naoko, Li, Jianying, Masai, Hisao, Vaziri, Cyrus
Formato: Online Artículo Texto
Lenguaje:English
Publicado: The American Society for Cell Biology 2012
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3350557/
https://www.ncbi.nlm.nih.gov/pubmed/22456510
http://dx.doi.org/10.1091/mbc.E11-10-0829
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author Barkley, Laura R.
Palle, Komaraiah
Durando, Michael
Day, Tovah A.
Gurkar, Aditi
Kakusho, Naoko
Li, Jianying
Masai, Hisao
Vaziri, Cyrus
author_facet Barkley, Laura R.
Palle, Komaraiah
Durando, Michael
Day, Tovah A.
Gurkar, Aditi
Kakusho, Naoko
Li, Jianying
Masai, Hisao
Vaziri, Cyrus
author_sort Barkley, Laura R.
collection PubMed
description The E3 ubiquitin ligase Rad18 chaperones DNA polymerase η (Polη) to sites of UV-induced DNA damage and monoubiquitinates proliferating cell nuclear antigen (PCNA), facilitating engagement of Polη 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 Polη-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 Polη association and did not redistribute Polη to nuclear foci or promote Polη−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.
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spelling pubmed-33505572012-07-30 c-Jun N-terminal kinase–mediated Rad18 phosphorylation facilitates Polη recruitment to stalled replication forks Barkley, Laura R. Palle, Komaraiah Durando, Michael Day, Tovah A. Gurkar, Aditi Kakusho, Naoko Li, Jianying Masai, Hisao Vaziri, Cyrus Mol Biol Cell Articles The E3 ubiquitin ligase Rad18 chaperones DNA polymerase η (Polη) to sites of UV-induced DNA damage and monoubiquitinates proliferating cell nuclear antigen (PCNA), facilitating engagement of Polη 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 Polη-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 Polη association and did not redistribute Polη to nuclear foci or promote Polη−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. The American Society for Cell Biology 2012-05-15 /pmc/articles/PMC3350557/ /pubmed/22456510 http://dx.doi.org/10.1091/mbc.E11-10-0829 Text en © 2012 Barkley et al. This article is distributed by The American Society for Cell Biology under license from the author(s). Two months after publication it is available to the public under an Attribution–Noncommercial–Share Alike 3.0 Unported Creative Commons License (http://creativecommons.org/licenses/by-nc-sa/3.0). “ASCB®,” “The American Society for Cell Biology®,” and “Molecular Biology of the Cell®” are registered trademarks of The American Society of Cell Biology.
spellingShingle Articles
Barkley, Laura R.
Palle, Komaraiah
Durando, Michael
Day, Tovah A.
Gurkar, Aditi
Kakusho, Naoko
Li, Jianying
Masai, Hisao
Vaziri, Cyrus
c-Jun N-terminal kinase–mediated Rad18 phosphorylation facilitates Polη recruitment to stalled replication forks
title c-Jun N-terminal kinase–mediated Rad18 phosphorylation facilitates Polη recruitment to stalled replication forks
title_full c-Jun N-terminal kinase–mediated Rad18 phosphorylation facilitates Polη recruitment to stalled replication forks
title_fullStr c-Jun N-terminal kinase–mediated Rad18 phosphorylation facilitates Polη recruitment to stalled replication forks
title_full_unstemmed c-Jun N-terminal kinase–mediated Rad18 phosphorylation facilitates Polη recruitment to stalled replication forks
title_short c-Jun N-terminal kinase–mediated Rad18 phosphorylation facilitates Polη recruitment to stalled replication forks
title_sort c-jun n-terminal kinase–mediated rad18 phosphorylation facilitates polη recruitment to stalled replication forks
topic Articles
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3350557/
https://www.ncbi.nlm.nih.gov/pubmed/22456510
http://dx.doi.org/10.1091/mbc.E11-10-0829
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