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Rad23 links DNA repair to the ubiquitin/proteasome pathway

Nature volume 391, pages 715–718 (1998)Cite this article

Abstract

Rad23 is an evolutionarily conserved protein that is important for nucleotide excision repair1,2,3. A regulatory role has been proposed for Rad23 because rad23 mutants are sensitive to ultraviolet light but are still capable of incising damaged DNA4,5. Here we show that Rad23 interacts with the 26S proteasome through an amino-terminal ubiquitin-like domain (UbLR23). The carboxy terminus of Rad23 binds to the Rad4 DNA repair protein and creates a link between the DNA repair and proteasome pathways. The ultraviolet sensitivity caused by deletion of the UbLR23 domain may therefore arise from its inability to interact with the proteasome. The fusion proteins glutathione S-transferase (GST)–Rad23 and Rad4–haemagglutinin (HA), and the proteasome subunits Cim3 and Cim5, cofractionate through consecutive chromatography steps. The ubiquitin-like domain of human Rad23 (UbLHRB) also interacts with the human proteasome. These results demonstrate that ubiquitin-like domains (UbLs) represent a new class of proteasome-interacting motifs.

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Figure 1: Rad23 interacts with the 26S proteasome.
Figure 2: Protease activity associated with Rad23.
Figure 3: Rad4–HA interacts with Rad23.
Figure 4: A complex of high Mr contains GST–Rad23, Rad4–HA and Cim5.
Figure 5: Rad23–HA is degraded by the proteasome.
Figure 6: Human HHR23-B interacts with Mss1.

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Acknowledgements

We thank C. Mann for antisera against Cim3/Sug1 and Cim5, M. Ghislain for cim3-1 and cim5-1 strains; J. Dohmen for the plasmid expressing Pre1-Flag; J. H. Hoeijmakers for cDNA clones expressing HHR23-A and HHR23-B; and D. Reinberg for HeLa cell extracts. This work was supported by a grant from the NIH (K.M.). I.V. was supported by a pre-doctoral fellowship from the NIH.

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Author notes

  1. Prasad Tongaonkar, Irving Vega, David Lambertson, Warren Potts and Kiran Madura: These authors contributed equallys to this work.

Authors and Affiliations

  1. Department of Biochemistry, Robert Wood Johnson Medical School, University of Medicine and Dentistry of New Jersey, 675 Hoes Lane, Piscataway, 08854, New Jersey, USA

    Cherylene Schauber, Li Chen, Prasad Tongaonkar, Irving Vega, David Lambertson, Warren Potts & Kiran Madura

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  1. Cherylene Schauber
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Correspondence to Kiran Madura.

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Schauber, C., Chen, L., Tongaonkar, P. et al. Rad23 links DNA repair to the ubiquitin/proteasome pathway. Nature 391, 715–718 (1998). https://doi.org/10.1038/35661

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