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A Rad26–Def1 complex coordinates repair and RNA pol II proteolysis in response to DNA damage

Nature volume 415, pages 929–933 (2002)Cite this article

Abstract

Eukaryotic cells use multiple, highly conserved mechanisms to contend with ultraviolet-light-induced DNA damage1. One important response mechanism is transcription-coupled repair (TCR), during which DNA lesions in the transcribed strand of an active gene are repaired much faster than in the genome overall2. In mammalian cells, defective TCR gives rise to the severe human disorder Cockayne's syndrome (CS)3. The best-studied CS gene, CSB, codes for a Swi/Snf-like DNA-dependent ATPase, whose yeast homologue is called Rad26 (ref. 4). Here we identify a yeast protein, termed Def1, which forms a complex with Rad26 in chromatin. The phenotypes of cells lacking DEF1 are consistent with a role for this factor in the DNA damage response, but Def1 is not required for TCR. Rather, def1 cells are compromised for transcript elongation, and are unable to degrade RNA polymerase II (RNAPII) in response to DNA damage. Our data suggest that RNAPII stalled at a DNA lesion triggers a coordinated rescue mechanism that requires the Rad26–Def1 complex, and that Def1 enables ubiquitination and proteolysis of RNAPII when the lesion cannot be rapidly removed by Rad26-promoted DNA repair.

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Figure 1: Affinity-purification of Rad26 and Def1.
Figure 2: UV-sensitivity of def1 strains.
Figure 3: The effect of DEF1 deletion on TCR and transcript elongation.
Figure 4: DEF1 is required for UV-induced ubiquitination and degradation of Rpb1.

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Acknowledgements

We thank the Cancer Research UK service facilities, especially fermentation services, for their help. We thank A. Grewal for assistance with peptide mass fingerprinting, and B. Winkler, P. Verrijzer, S. West and T. Lindahl for comments on the manuscript. This work was supported by grants from the ICRF and the Human Frontier Science Programme to J.Q.S., and by a NCI Core Grant to P.T.

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Authors and Affiliations

  1. Mechanisms of Gene Transcription Laboratory, Cancer Research UK, Clare Hall Laboratories, Blanche Lane, South Mimms, EN6 3LD, Hertfordshire, UK

    Elies C. Woudstra, Chris Gilbert, Jane Fellows & Jesper Q. Svejstrup

  2. MGC Department of Molecular Genetics, Leiden Institute of Chemistry, Leiden University, Leiden, 2300 RA, The Netherlands

    Lars Jansen & Jaap Brouwer

  3. Molecular Biology Programme, Memorial Sload-Kettering Cancer Center, York Avenue 1275, New York, 10021, USA

    Hediye Erdjument-Bromage & Paul Tempst

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  1. Elies C. Woudstra
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Correspondence to Jesper Q. Svejstrup.

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Woudstra, E., Gilbert, C., Fellows, J. et al. A Rad26–Def1 complex coordinates repair and RNA pol II proteolysis in response to DNA damage. Nature 415, 929–933 (2002). https://doi.org/10.1038/415929a

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