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Regulation of monoubiquitinated PCNA by DUB autocleavage

Nature Cell Biology volume 8, pages 341–347 (2006)Cite this article

An Erratum to this article was published on 01 April 2006

Abstract

Monoubiquitination is a reversible post-translational protein modification that has an important regulatory function in many biological processes, including DNA repair. Deubiquitinating enzymes (DUBs) are proteases that are negative regulators of monoubiquitination, but little is known about their regulation and contribution to the control of conjugated-substrate levels. Here, we show that the DUB ubiquitin specific protease 1 (USP1) deubiquitinates the DNA replication processivity factor, PCNA, as a safeguard against error-prone translesion synthesis (TLS) of DNA. Ultraviolet (UV) irradiation inactivates USP1 through an autocleavage event, thus enabling monoubiquitinated PCNA to accumulate and to activate TLS. Significantly, the site of USP1 cleavage is immediately after a conserved internal ubiquitin-like diglycine (Gly–Gly) motif. This mechanism is reminiscent of the processing of precursors of ubiquitin and ubiquitin-like modifiers by DUBs. Our results define a regulatory mechanism for protein ubiquitination that involves the signal-induced degradation of an inhibitory DUB.

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Figure 1: Knockdown of USP1 increases PCNA monoubiquitination.
Figure 2: UV damage degrades USP1 and increases PCNA monoubiquitination.
Figure 3: Inhibition of diverse DNA repair pathways does not affect UV-induced USP1 degradation.
Figure 4: Degradation of USP1 requires its own catalytic activity.
Figure 5: The conserved diglycine motif of USP1 is required for its autocleavage.
Figure 6: Increased mutation frequency in cells depleted of USP1.

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Acknowledgements

We thank D. Finley and members of the D'Andrea laboratory for critical reading of the manuscript. We are grateful to V. Notenboom for technical assistance. We thank T. Taniguchi for RAD18 and PCNA cDNAs and A. Lehmann for GFP-polη expression constructs. We are grateful to M. M. Seidman for generously providing the pSP189 plasmid and the MBM7070 bacterial strain. This work was supported by grants from the National Institutes of Health (NIH, A.D.D.) and was funded in part by the Doris Duke Charitable Foundation (A.D.D.). T.T.H. is a Blount fellow of the Damon Runyon Cancer Research Foundation.

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Authors and Affiliations

  1. Department of Radiation Oncology, Dana-Farber Cancer Institute, Harvard Medical School, Boston, 02115, MA, USA

    Tony T. Huang, Kanchan D. Mirchandani, Martin A. Cohn & Alan D. D'Andrea

  2. Division of Molecular Carcinogenesis and Center for Biomedical Genetics, The Netherlands Cancer Institute, Amsterdam, The Netherlands

    Sebastian M.B. Nijman & René Bernards

  3. Department of Pathology, Harvard Medical School, Boston, 02115, MA, USA

    Paul J. Galardy & Hidde L. Ploegh

  4. Department of Cell Biology, Harvard Medical School, Boston, 02115, MA, USA

    Wilhelm Haas & Steven P. Gygi

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Huang, T., Nijman, S., Mirchandani, K. et al. Regulation of monoubiquitinated PCNA by DUB autocleavage. Nat Cell Biol 8, 341–347 (2006). https://doi.org/10.1038/ncb1378

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