PLoS ONE (Jan 2017)

Structural basis for the substrate selectivity of Helicobacter pylori NucT nuclease activity.

  • Louisa Celma,
  • Christopher Corbinais,
  • Julien Vercruyssen,
  • Xavier Veaute,
  • Inès Li de la Sierra-Gallay,
  • Raphaël Guérois,
  • Didier Busso,
  • Aurélie Mathieu,
  • Stéphanie Marsin,
  • Sophie Quevillon-Cheruel,
  • J Pablo Radicella

DOI
https://doi.org/10.1371/journal.pone.0189049
Journal volume & issue
Vol. 12, no. 12
p. e0189049

Abstract

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The Phospholipase D (PLD) superfamily of proteins includes a group of enzymes with nuclease activity on various nucleic acid substrates. Here, with the aim of better understanding the substrate specificity determinants in this subfamily, we have characterised the enzymatic activity and the crystal structure of NucT, a nuclease implicated in Helicobacter pylori purine salvage and natural transformation and compared them to those of its bacterial and mammalian homologues. NucT exhibits an endonuclease activity with a strong preference for single stranded nucleic acids substrates. We identified histidine124 as essential for the catalytic activity of the protein. Comparison of the NucT crystal structure at 1.58 Å resolution reported here with those of other members of the sub-family suggests that the specificity of NucT for single-stranded nucleic acids is provided by the width of a positively charged groove giving access to the catalytic site.