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The crystal structure of flap endonuclease-1 from Methanococcus jannaschii

Nature Structural Biology volume 5, pages 707–713 (1998)Cite this article

Abstract

Flap endonuclease-1 (FEN-1), a structure specific nuclease, is an essential enzyme for eukaryotic DNA replication and repair. The crystal structure of FEN-1 from Methanococcus jannaschii, determined at 2.0 Å resolution, reveals an active site with two metal ions residing on top of a deep cleft where several conserved acidic residues are clustered. Near the active site, a long flexible loop comprised of many basic and aromatic residues forms a hole large enough to accommodate the DNA substrate. Deletion mutations in this loop significantly decreased the nuclease activity and specificity of FEN-1, suggesting that the loop is critical for recognition and cleavage of the junction between single and double-stranded regions of flap DNA.

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Figure 1: a, Multiple isomorphous replacement (MIR) electron density map of Mj FEN-1 at 3.0 Å resolution.
Figure 2: Stereo diagrams showing the significant interactions within Mj FEN-1.
Figure 3: a, The endonuclease activities of Mj FEN-1 for 5' flap (lanes 2–6) and pseudo Y DNA (lanes 7–11).
Figure 4: a, A stereo diagram showing the solvent accessible surface of Mj FEN-1.

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Acknowledgements

We are grateful to M. S. Park (Los Alamos National Lab), S. S. Kim, E. Kim (L.G. Biotech) for their invaluable comments. We thanks to K. W. Bae and C. S. Cho for technical assistance. This work was supported by Biotech 2000 program from MOST and KIST 2000 program from KIST.

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    1. Structural Biology Center, Korea Institute of Science & Technology, P.O. Box 13, Cheongryang, Seoul, 130-650, South Korea

      Kwang Yeon Hwang, Kyuwon Baek, Hye-Yeon Kim & Yunje Cho

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    Correspondence to Yunje Cho.

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    Hwang, K., Baek, K., Kim, HY. et al. The crystal structure of flap endonuclease-1 from Methanococcus jannaschii . Nat Struct Mol Biol 5, 707–713 (1998). https://doi.org/10.1038/1406

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