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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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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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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DOI: https://doi.org/10.1038/1406
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