Abstract

The dUTPase is a key DNA repair enzyme in Mycobacterium tuberculosis, and it may serve as a novel promising anti-tuberculosis target. Stl repressor from Staphylococcus aureus was shown to bind to and inhibit dUTPases from various sources, and its expression in mycobacterial cells interfered with cell growth. To fine-tune and optimize Stl-induced inhibition of mycobacterial dUTPase, we aimed to decipher the molecular details of this interaction. Structural background of the complex between dUTPase and a truncated Stl lacking the repressor C-terminal homodimerization domain has been described, however, the effects of this truncation of Stl on enzyme binding and inhibition are still not known. Using several independent biophysical, structural and enzyme kinetic methods, here we show that lack of the repressor homodimerization domain strongly perturbs both enzyme binding and inhibition. We also investigated the role of a mycobacteria-specific loop in the Stl-interaction. Our results show that removal of this loop leads to a ten-fold increase in the apparent inhibition constant of Stl. We present a high-resolution three-dimensional structure of mycobacterial dUTPase lacking the genus-specific loop for structural insight. Our present data suggest that potent inhibition of mycobacterial dUTPase by Stl requires the wild-type full-length protein context.

Details

Title
The homodimerization domain of the Stl repressor is crucial for efficient inhibition of mycobacterial dUTPase
Author
Tóth, Zoé S. 1 ; Leveles, Ibolya 2 ; Nyíri, Kinga 2 ; Nagy, Gergely N. 2 ; Harmat, Veronika 3 ; Jaroentomeechai, Thapakorn 4 ; Ozohanics, Oliver 5 ; Miller, Rebecca L. 4 ; Álvarez, Marina Ballesteros 6 ; Vértessy, Beáta G. 2 ; Benedek, András 2 

 HUN-REN Research Centre for Natural Sciences, Institute of Molecular Life Sciences, Budapest, Hungary (GRID:grid.425578.9) (ISNI:0000 0004 0512 3755); Budapest University of Technology and Economics, Department of Applied Biotechnology and Food Science, Faculty of Chemical Technology and Biotechnology, Budapest, Hungary (GRID:grid.6759.d) (ISNI:0000 0001 2180 0451); ELTE Eötvös Loránd University, Doctoral School of Biology, Institute of Biology, Budapest, Hungary (GRID:grid.5591.8) (ISNI:0000 0001 2294 6276) 
 HUN-REN Research Centre for Natural Sciences, Institute of Molecular Life Sciences, Budapest, Hungary (GRID:grid.425578.9) (ISNI:0000 0004 0512 3755); Budapest University of Technology and Economics, Department of Applied Biotechnology and Food Science, Faculty of Chemical Technology and Biotechnology, Budapest, Hungary (GRID:grid.6759.d) (ISNI:0000 0001 2180 0451) 
 ELTE Eötvös Loránd University, Laboratory of Structural Chemistry and Biology, Institute of Chemistry, Budapest, Hungary (GRID:grid.5591.8) (ISNI:0000 0001 2294 6276); HUN-REN-ELTE Protein Modelling Research Group, Hungarian Research Network, Budapest, Hungary (GRID:grid.5591.8) (ISNI:0000 0004 9284 0620) 
 University of Copenhagen, Copenhagen Center for Glycomics, Department of Cellular and Molecular Medicine, Copenhagen, Denmark (GRID:grid.5254.6) (ISNI:0000 0001 0674 042X) 
 Semmelweis University, Department of Medical Biochemistry, Budapest, Hungary (GRID:grid.11804.3c) (ISNI:0000 0001 0942 9821) 
 Budapest University of Technology and Economics, Department of Applied Biotechnology and Food Science, Faculty of Chemical Technology and Biotechnology, Budapest, Hungary (GRID:grid.6759.d) (ISNI:0000 0001 2180 0451) 
Pages
27171
Publication year
2024
Publication date
2024
Publisher
Nature Publishing Group
e-ISSN
20452322
Source type
Scholarly Journal
Language of publication
English
ProQuest document ID
3125884478
Copyright
© The Author(s) 2024. This work is published under http://creativecommons.org/licenses/by-nc-nd/4.0/ (the “License”). Notwithstanding the ProQuest Terms and Conditions, you may use this content in accordance with the terms of the License.