Direct contacts between conserved motifs of different subunits provide major contribution to active site organization in human and mycobacterial dUTPases

Eniko Takács, Gergely Nagy, Ibolya Leveles, Veronika Harmat, Anna Lopata, Judit Tóth, Beáta G. Vértessy

Research output: Contribution to journalArticle

12 Citations (Scopus)

Abstract

dUTP pyrophosphatases (dUTPases) are essential for genome integrity. Recent results allowed characterization of the role of conserved residues. Here we analyzed the Asp/Asn mutation within conserved Motif I of human and mycobacterial dUTPases, wherein the Asp residue was previously implicated in Mg2+-coordination. Our results on transient/steady-state kinetics, ligand binding and a 1.80Å resolution structure of the mutant mycobacterial enzyme, in comparison with wild type and C-terminally truncated structures, argue that this residue has a major role in providing intra- and intersubunit contacts, but is not essential for Mg2+ accommodation. We conclude that in addition to the role of conserved motifs in substrate accommodation, direct subunit interaction between protein atoms of active site residues from different conserved motifs are crucial for enzyme function.

Original languageEnglish
Pages (from-to)3047-3054
Number of pages8
JournalFEBS letters
Volume584
Issue number14
DOIs
Publication statusPublished - Jul 2010

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Keywords

  • Asparagine
  • Aspartate
  • DNA repair
  • DUTPase
  • Mycobacterium tuberculosis
  • Nucleotide hydrolysis
  • Oligomer

ASJC Scopus subject areas

  • Biophysics
  • Structural Biology
  • Biochemistry
  • Molecular Biology
  • Genetics
  • Cell Biology

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