Direct regulation of Chk1 protein stability by E3 ubiquitin ligase HUWE1

Katelyn B. Cassidy; Scott Bang; Manabu Kurokawa; Scott A. Gerber

doi:10.1111/febs.15132

journal article Nov 29, 2019

Direct regulation of Chk1 protein stability by E3 ubiquitin ligase HUWE1

Katelyn B. Cassidy Scott Bang Manabu Kurokawa Scott A. Gerber

The FEBS Journal Vol. 287 No. 10 pp. 1985-1999 · Wiley

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Abstract

The HECT E3 ubiquitin ligase HUWE1 is required for a wide array of important functions in cell biology. Although HUWE1 is known to play a role in DNA damage signaling, the mechanism(s) that underlie this function remain elusive. HUWE1 regulates effectors of DNA replication and genotoxic stress tolerance. However, the loss of HUWE1 can also result in the accrual of significant endogenous DNA damage due to insufficient remediation of replication stress induced by an overabundance of key substrates. We discovered that HUWE1 depletion leads to a significant increase in levels of the single‐strand break effector kinase Chk1, independent of the DNA damage response, activation of apical DNA damage repair (DDR) signaling kinases (ATM and ATR), and the tumor suppressor p53. We also identified multiple lysine residues on Chk1 that are polyubiquitinated by HUWE1 in vitro, many of which are within the kinase domain. HUWE1 knockdown also markedly prolonged the protein half‐life of Chk1 in steady‐state conditions and resulted in greater stabilization of Chk1 protein than depletion of Cul4A, an E3 ubiquitin ligase previously described to control Chk1 abundance. Moreover, prolonged replication stress induced by hydroxyurea or camptothecin resulted in a reduction of Chk1 protein levels, which was rescued by HUWE1 knockdown. Our study indicates that HUWE1 plays a significant role in the regulation of the DDR signaling pathway by directly modulating the abundance of Chk1 protein.

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Details

Published: Nov 29, 2019
Vol/Issue: 287(10)
Pages: 1985-1999
License: View

Authors

K

Katelyn B. Cassidy

Department of Molecular & Systems Biology Geisel School of Medicine Hanover NH USA

S

Scott Bang

Department of Biological Sciences Kent State University Kent OH USA

M

Manabu Kurokawa

Department of Molecular & Systems Biology Geisel School of Medicine Hanover NH USA; Department of Biological Sciences Kent State University Kent OH USA; Norris Cotton Cancer Center Geisel School of Medicine Lebanon NH USA

S

Scott A. Gerber

Department of Molecular & Systems Biology Geisel School of Medicine Hanover NH USA; Norris Cotton Cancer Center Geisel School of Medicine Lebanon NH USA

Funding

National Cancer Institute Award: R01CA155260

National Institute of General Medical Sciences Award: R01GM122846

American Cancer Society Award: IRG‐82‐003‐30

Cite This Article

Katelyn B. Cassidy, Scott Bang, Manabu Kurokawa, et al. (2019). Direct regulation of Chk1 protein stability by E3 ubiquitin ligase HUWE1. The FEBS Journal, 287(10), 1985-1999. https://doi.org/10.1111/febs.15132

Direct regulation of Chk1 protein stability by E3 ubiquitin ligase HUWE1

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