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Methylation of FEN1 suppresses nearby phosphorylation and facilitates PCNA binding

Nature Chemical Biology volume 6, pages 766–773 (2010)Cite this article

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Abstract

Flap endonuclease 1 (FEN1), a structure-specific endo- and exonuclease, has multiple functions that determine essential biological processes, such as cell proliferation and cell death. As such, the enzyme must be precisely regulated to execute each of its functions with the right timing and in a specific subcellular location. Here we report that FEN1 is methylated at arginine residues, primarily at Arg192. The methylation suppresses FEN1 phosphorylation at Ser187. The methylated form, but not the phosphorylated form, of FEN1 strongly interacts with proliferating cell nuclear antigen (PCNA), ensuring the 'on' and 'off' timing of its reaction. Mutations of FEN1 disrupting arginine methylation and PCNA interaction result in unscheduled phosphorylation and a failure to localize to DNA replication or repair foci. This consequently leads to a defect in Okazaki fragment maturation, a delay in cell cycle progression, impairment of DNA repair and a high frequency of genome-wide mutations.

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Figure 1: In vivo and in vitro methylation of FEN1 by PRMT5.
Figure 2: Identification and validation of FEN1 methylation sites.
Figure 3: Methylation of FEN1 at Arg192 inhibits its phosphorylation at Ser187.
Figure 4: Methylation of FEN1 ensures its interaction with PCNA via suppression of its phosphorylation.
Figure 5: Methylation defect retards Okazaki fragment maturation and DNA replication and induces DNA damage.
Figure 6: Methylation of FEN1 is induced by H2O2 and is important for DNA repair.

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Acknowledgements

We thank G. Tsaprailis in the Proteomics Core facility of University of Arizona for technical assistance in determining the methylation sites on FEN1. We thank S.R. da Costa for editorial assistance. This work was supported by US National Institutes of Health grants RO1 CA073764 and R01CA085344 to B.S.

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Authors and Affiliations

  1. Department of Cancer Biology, City of Hope National Medical Center and Beckman Research Institute, Duarte, California, USA

    Zhigang Guo, Li Zheng, Hong Xu, Huifang Dai, Mian Zhou, Mary Rose Pascua & Binghui Shen

  2. Department of Pharmacology, University of Arizona, Tucson, Arizona, USA

    Qin M Chen

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Contributions

Z.G., L.Z. and B.S. designed the experiments, analyzed the data and wrote the manuscript. H.X. purified recombinant proteins. H.D., M.Z. and M.R.P. performed cell biological experiments. Q.M.C. performed the experiments with mass spectrometry and analyzed the data.

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Correspondence to Binghui Shen.

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Guo, Z., Zheng, L., Xu, H. et al. Methylation of FEN1 suppresses nearby phosphorylation and facilitates PCNA binding. Nat Chem Biol 6, 766–773 (2010). https://doi.org/10.1038/nchembio.422

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