Cargando…
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...
Autores principales: | , , , , , , , , |
---|---|
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 |
_version_ | 1782232689079746560 |
---|---|
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. |
format | Online Article Text |
id | pubmed-3350557 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2012 |
publisher | The American Society for Cell Biology |
record_format | MEDLINE/PubMed |
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 |
work_keys_str_mv | AT barkleylaurar cjunnterminalkinasemediatedrad18phosphorylationfacilitatespolērecruitmenttostalledreplicationforks AT pallekomaraiah cjunnterminalkinasemediatedrad18phosphorylationfacilitatespolērecruitmenttostalledreplicationforks AT durandomichael cjunnterminalkinasemediatedrad18phosphorylationfacilitatespolērecruitmenttostalledreplicationforks AT daytovaha cjunnterminalkinasemediatedrad18phosphorylationfacilitatespolērecruitmenttostalledreplicationforks AT gurkaraditi cjunnterminalkinasemediatedrad18phosphorylationfacilitatespolērecruitmenttostalledreplicationforks AT kakushonaoko cjunnterminalkinasemediatedrad18phosphorylationfacilitatespolērecruitmenttostalledreplicationforks AT lijianying cjunnterminalkinasemediatedrad18phosphorylationfacilitatespolērecruitmenttostalledreplicationforks AT masaihisao cjunnterminalkinasemediatedrad18phosphorylationfacilitatespolērecruitmenttostalledreplicationforks AT vaziricyrus cjunnterminalkinasemediatedrad18phosphorylationfacilitatespolērecruitmenttostalledreplicationforks |