Structural model of human dUTPase in complex with a novel proteinaceous inhibitor

Kinga Nyíri, Haydyn D.T. Mertens, Borbála Tihanyi, Gergely N. Nagy, Bianka Kohegyi, Judit Matejka, Matthew J. Harris, Judit E. Szabó, Veronika Papp-Kádár, Veronika Németh-Pongrácz, Olivér Ozohanics, Károly Vékey, Dmitri I. Svergun, Antoni J. Borysik, Beáta G. Vértessy

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Abstract

Human deoxyuridine 5′-triphosphate nucleotidohydrolase (dUTPase), essential for DNA integrity, acts as a survival factor for tumor cells and is a target for cancer chemotherapy. Here we report that the Staphylococcal repressor protein StlSaPIBov1 (Stl) forms strong complex with human dUTPase. Functional analysis reveals that this interaction results in significant reduction of both dUTPase enzymatic activity and DNA binding capability of Stl. We conducted structural studies to understand the mechanism of this mutual inhibition. Small-angle X-ray scattering (SAXS) complemented with hydrogen-deuterium exchange mass spectrometry (HDX-MS) data allowed us to obtain 3D structural models comprising a trimeric dUTPase complexed with separate Stl monomers. These models thus reveal that upon dUTPase-Stl complex formation the functional homodimer of Stl repressor dissociates, which abolishes the DNA binding ability of the protein. Active site forming dUTPase segments were directly identified to be involved in the dUTPase-Stl interaction by HDX-MS, explaining the loss of dUTPase activity upon complexation. Our results provide key novel structural insights that pave the way for further applications of the first potent proteinaceous inhibitor of human dUTPase.

Original languageEnglish
Article number4326
JournalScientific reports
Volume8
Issue number1
DOIs
Publication statusPublished - Dec 1 2018

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Nyíri, K., Mertens, H. D. T., Tihanyi, B., Nagy, G. N., Kohegyi, B., Matejka, J., Harris, M. J., Szabó, J. E., Papp-Kádár, V., Németh-Pongrácz, V., Ozohanics, O., Vékey, K., Svergun, D. I., Borysik, A. J., & Vértessy, B. G. (2018). Structural model of human dUTPase in complex with a novel proteinaceous inhibitor. Scientific reports, 8(1), [4326]. https://doi.org/10.1038/s41598-018-22145-8