TY - JOUR
T1 - HUWE1 interacts with PCNA to alleviate replication stress
AU - Choe, Katherine N.
AU - Nicolae, Claudia M.
AU - Constantin, Daniel
AU - Imamura Kawasawa, Yuka
AU - Delgado-Diaz, Maria Rocio
AU - De, Subhajyoti
AU - Freire, Raimundo
AU - Smits, Veronique A.
AU - Moldovan, George Lucian
N1 - Funding Information:
We would like to thank Sanziana Rotariu and Hejuan He for early contributions to this work, and Alan D'Andrea, James Broach, Stefan Jentsch, Wafik El Deiry, David Cortez, Wojciech Piwko, Kyungjae Myung, David Kozono, Sergei Grigoryev, Kristin Eckert, Thomas Spratt, Faoud Ishmael, Laura Carrel, and Gregory Yochum for materials, support, and advice. This work was supported by: NIH 1R01ES026184, Department of Defense CA140303, St. Baldrick Foundation, V Foundation, Concern Foundation, Gittlen Foundation (to GLM); American Cancer Society ACS IRG 13 043 01 (to CMN); American Cancer Society ACS IRG 57 001 53, Lung Cancer Colorado Fund, and United Against Lung Cancer (to SD)
Publisher Copyright:
© 2016 The Authors.
PY - 2016/6/1
Y1 - 2016/6/1
N2 - Defects in DNA replication, DNA damage response, and DNA repair compromise genomic stability and promote cancer development. In particular, unrepaired DNA lesions can arrest the progression of the DNA replication machinery during S-phase, causing replication stress, mutations, and DNA breaks. HUWE1 is a HECT-type ubiquitin ligase that targets proteins involved in cell fate, survival, and differentiation. Here, we report that HUWE1 is essential for genomic stability, by promoting replication of damaged DNA. We show that HUWE1-knockout cells are unable to mitigate replication stress, resulting in replication defects and DNA breakage. Importantly, we find that this novel role of HUWE1 requires its interaction with the replication factor PCNA, a master regulator of replication fork restart, at stalled replication forks. Finally, we provide evidence that HUWE1 mono-ubiquitinates H2AX to promote signaling at stalled forks. Altogether, our work identifies HUWE1 as a novel regulator of the replication stress response. Synopsis The HUWE1 ubiquitin ligase is recruited to stalled replication forks by the replication factor PCNA. HUWE1 ubiquitinates H2AX to promote restart of stalled replication forks and mitigate replication stress. HUWE1 contains a PCNA interacting domain (PIP-box) required for PCNA interaction. HUWE1-knockout cells show increased replication stress, which can be corrected by re-expression of wild-type but not PCNA interaction-deficient HUWE1 mutant. HUWE1 mono-ubiquitinates H2AX to promote γH2AX signaling, recruitment of repair proteins, and restart of stalled replication forks. The HUWE1 ubiquitin ligase is recruited to stalled replication forks by the replication factor PCNA. HUWE1 ubiquitinates H2AX to promote restart of stalled replication forks and mitigate replication stress.
AB - Defects in DNA replication, DNA damage response, and DNA repair compromise genomic stability and promote cancer development. In particular, unrepaired DNA lesions can arrest the progression of the DNA replication machinery during S-phase, causing replication stress, mutations, and DNA breaks. HUWE1 is a HECT-type ubiquitin ligase that targets proteins involved in cell fate, survival, and differentiation. Here, we report that HUWE1 is essential for genomic stability, by promoting replication of damaged DNA. We show that HUWE1-knockout cells are unable to mitigate replication stress, resulting in replication defects and DNA breakage. Importantly, we find that this novel role of HUWE1 requires its interaction with the replication factor PCNA, a master regulator of replication fork restart, at stalled replication forks. Finally, we provide evidence that HUWE1 mono-ubiquitinates H2AX to promote signaling at stalled forks. Altogether, our work identifies HUWE1 as a novel regulator of the replication stress response. Synopsis The HUWE1 ubiquitin ligase is recruited to stalled replication forks by the replication factor PCNA. HUWE1 ubiquitinates H2AX to promote restart of stalled replication forks and mitigate replication stress. HUWE1 contains a PCNA interacting domain (PIP-box) required for PCNA interaction. HUWE1-knockout cells show increased replication stress, which can be corrected by re-expression of wild-type but not PCNA interaction-deficient HUWE1 mutant. HUWE1 mono-ubiquitinates H2AX to promote γH2AX signaling, recruitment of repair proteins, and restart of stalled replication forks. The HUWE1 ubiquitin ligase is recruited to stalled replication forks by the replication factor PCNA. HUWE1 ubiquitinates H2AX to promote restart of stalled replication forks and mitigate replication stress.
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U2 - 10.15252/embr.201541685
DO - 10.15252/embr.201541685
M3 - Article
C2 - 27146073
AN - SCOPUS:84971499032
SN - 1469-221X
VL - 17
SP - 874
EP - 886
JO - EMBO Reports
JF - EMBO Reports
IS - 6
ER -