The role of a key amino acid position in species- specific proteinaceous dUTPase inhibition

András Benedek, Fanni Temesváry-Kis, Tamjidmaa Khatanbaatar, Ibolya Leveles, Éva Viola Surányi, Judit Eszter Szabó, Lívius Wunderlich, Beáta G. Vértessy

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1 Citation (Scopus)

Abstract

Protein inhibitors of key DNA repair enzymes play an important role in deciphering physiological pathways responsible for genome integrity, and may also be exploited in biomedical research. The staphylococcal repressor StlSaPIbov1 protein was described to be an efficient inhibitor of dUTPase homologues showing a certain degree of species-specificity. In order to provide insight into the inhibition mechanism, in the present study we investigated the interaction of StlSaPIbov1 and Escherichia coli dUTPase. Although we observed a strong interaction of these proteins, unexpectedly the E. coli dUTPase was not inhibited. Seeking a structural explanation for this phenomenon, we identified a key amino acid position where specific mutations sensitized E. coli dUTPase to StlSaPIbov1 inhibition. We solved the three-dimensional (3D) crystal structure of such a mutant in complex with the substrate analogue dUPNPP and surprisingly found that the Cterminal arm of the enzyme, containing the P-loop-like motif was ordered in the structure. This segment was never localized before in any other E. coli dUTPase crystal structures. The 3D structure in agreement with solution phase experiments suggested that ordering of the flexible C-terminal segment upon substrate binding is a major factor in defining the sensitivity of E. coli dUTPase for StlSaPIbov1 inhibition.

Original languageEnglish
Article number221
JournalBiomolecules
Volume9
Issue number6
DOIs
Publication statusPublished - Jun 2019

Keywords

  • C-terminal arm
  • Crystal structure
  • DUTPase
  • Inhibition
  • StlSaPIbov1
  • Trimer

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology

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    Benedek, A., Temesváry-Kis, F., Khatanbaatar, T., Leveles, I., Surányi, É. V., Szabó, J. E., Wunderlich, L., & Vértessy, B. G. (2019). The role of a key amino acid position in species- specific proteinaceous dUTPase inhibition. Biomolecules, 9(6), [221]. https://doi.org/10.3390/biom9060221