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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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.
Reprints and permissions information is available at npg.nature.com/reprintsandpermissions. The authors declare no competing financial interests.
Additional methods and Supplementary Table 1 showing yeast strains used in this study. (PDF 59 kb)
Legends to accompany the Supplementary Figures S1-S5. (PDF 35 kb)
PCNA ubiquitination is induced by DNA damage specifically in S-Phase. (PDF 79 kb)
Survival rates after ultraviolet irradiation. (PDF 138 kb)
PCNA SUMOylation levels are influenced by Srs2. (PDF 163 kb)
Survival rates in response to ultraviolet radiation. (PDF 91 kb)
The PCNA-SUMOSrs2 check operates in diploid cells. (PDF 231 kb)
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Pfander, B., Moldovan, G., Sacher, M. et al. SUMO-modified PCNA recruits Srs2 to prevent recombination during S phase. Nature 436, 428–433 (2005) doi:10.1038/nature03665
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