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SUMO-modified PCNA recruits Srs2 to prevent recombination during S phase

Nature volume 436, pages 428–433 (2005)Cite this article

Abstract

Damaged DNA, if not repaired before replication, can lead to replication fork stalling and genomic instability1,2,3; however, cells can switch to different damage bypass modes that permit replication across lesions. Two main bypasses are controlled by ubiquitin modification of proliferating cell nuclear antigen (PCNA), a homotrimeric DNA-encircling protein that functions as a polymerase processivity factor and regulator of replication-linked functions4,5. Upon DNA damage, PCNA is modified at the conserved lysine residue 164 by either mono-ubiquitin or a lysine-63-linked multi-ubiquitin chain5, which induce error-prone or error-free replication bypasses of the lesions5,6. In S phase, even in the absence of exogenous DNA damage, yeast PCNA can be alternatively modified by the small ubiquitin-related modifier protein SUMO5; however the consequences of this remain controversial5,6,7. Here we show by genetic analysis that SUMO-modified PCNA functionally cooperates with Srs2, a helicase that blocks recombinational repair by disrupting Rad51 nucleoprotein filaments8,9. Moreover, Srs2 displays a preference for interacting directly with the SUMO-modified form of PCNA, owing to a specific binding site in its carboxy-terminal tail. Our finding suggests a model in which SUMO-modified PCNA recruits Srs2 in S phase in order to prevent unwanted recombination events of replicating chromosomes.

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Figure 1: Suppression of DNA damage sensitivity of RAD6 pathway mutants by deficiency in PCNA SUMOylation requires homologous recombination.
Figure 2: Srs2 preferentially binds SUMOylated PCNA.
Figure 3: Mutants deficient in PCNA SUMOylation and in the C-terminal tail of Srs2 ( srs2ΔC ) are epistatic with respect to suppression of RAD6 pathway mutants.
Figure 4: Influence of the PCNA–SUMO–Srs2 check on recombination and mutator phenotypes.

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Acknowledgements

We thank U. Cramer for technical assistance, S. Kumar for the gift of the SRS2ΔN clone, D. Siepe for computational analysis, and E. S. Johnson, H. L. Klein, C. Pohl, H. Richly and H. D. Ulrich for materials. This work is supported (to S. J.) by the Max Planck Society, Deutsche Krebshilfe, Deutsche Forschungsgemeinschaft, and Fonds der chemischen Industrie.

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  1. Carsten Hoege

    Present address: Max Planck Institute of Molecular Cell Biology and Genetics, Pfotenhauerstrasse 108, 01307, Dresden, Germany

Authors and Affiliations

  1. Department of Molecular Cell Biology, Max Planck Institute of Biochemistry, Am Klopferspitz 18, 82152, Martinsried, Germany

    Boris Pfander, George-Lucian Moldovan, Meik Sacher, Carsten Hoege & Stefan Jentsch

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Correspondence to Stefan Jentsch.

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Supplementary information

Supplementary Methods

Additional methods and Supplementary Table 1 showing yeast strains used in this study. (PDF 59 kb)

Supplementary Figure Legends

Legends to accompany the Supplementary Figures S1-S5. (PDF 35 kb)

Supplementary Figure S1

PCNA ubiquitination is induced by DNA damage specifically in S-Phase. (PDF 79 kb)

Supplementary Figure S2

Survival rates after ultraviolet irradiation. (PDF 138 kb)

Supplementary Figure S3

PCNA SUMOylation levels are influenced by Srs2. (PDF 163 kb)

Supplementary Figure S4

Survival rates in response to ultraviolet radiation. (PDF 91 kb)

Supplementary Figure S5

The PCNA-SUMOSrs2 check operates in diploid cells. (PDF 231 kb)

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Pfander, B., Moldovan, GL., Sacher, M. et al. SUMO-modified PCNA recruits Srs2 to prevent recombination during S phase. Nature 436, 428–433 (2005). https://doi.org/10.1038/nature03665

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