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A novel endonuclease mechanism directly visualized for I-PpoI.

Nat Struct Biol 6 1096-9 (1999)
Cited: 68 times
EuropePMC logo PMID: 10581547

Abstract

A novel mechanism of DNA endonucleolytic cleavage has been visualized for the homing endonuclease I-PpoI by trapping the uncleaved enzyme-substrate complex and comparing it to the previously visualized product complex. This enzyme employs a unique single metal mechanism. A magnesium ion is coordinated by an asparagine residue and two DNA oxygen atoms and stabilizes the phosphoanion transition state and the 3'oxygen leaving group. A hydrolytic water molecule is activated by a histidine residue for an in-line attack on the scissile phosphate. A strained enzyme-substrate-metal complex is formed before cleavage, then relaxed during the reaction.

Reviews citing this publication (10)

  1. Structure and function of type II restriction endonucleases. Pingoud A, Jeltsch A. Nucleic Acids Res 29 3705-3727 (2001)
  2. Homing endonucleases: structural and functional insight into the catalysts of intron/intein mobility. Chevalier BS, Stoddard BL. Nucleic Acids Res 29 3757-3774 (2001)
  3. Type II restriction endonucleases--a historical perspective and more. Pingoud A, Wilson GG, Wende W. Nucleic Acids Res 42 7489-7527 (2014)
  4. Structure and function of nucleases in DNA repair: shape, grip and blade of the DNA scissors. Nishino T, Morikawa K. Oncogene 21 9022-9032 (2002)
  5. Homing endonucleases: from basics to therapeutic applications. Marcaida MJ, Muñoz IG, Blanco FJ, Prieto J, Montoya G. Cell Mol Life Sci 67 727-748 (2010)
  6. Roles of metal ions in nucleases. Dupureur CM. Curr Opin Chem Biol 12 250-255 (2008)
  7. Restriction endonucleases: classification, properties, and applications. Williams RJ. Mol Biotechnol 23 225-243 (2003)
  8. One is enough: insights into the two-metal ion nuclease mechanism from global analysis and computational studies. Dupureur CM. Metallomics 2 609-620 (2010)
  9. The use of divalent metal ions by type II topoisomerases. Deweese JE, Osheroff N. Metallomics 2 450-459 (2010)
  10. Structural, functional and evolutionary relationships between homing endonucleases and proteins from their host organisms. Taylor GK, Stoddard BL. Nucleic Acids Res 40 5189-5200 (2012)

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