title: Cross-species inhibition of dUTPase via the Staphylococcal Stl protein perturbs dNTP pool and colony formation in Mycobacterium.
abstract: Proteins responsible for the integrity of the genome are often used targets in drug therapies against various diseases. The inhibitors of these proteins are also important to study the pathways in genome integrity maintenance. A prominent example is Ugi, a well known cross-species inhibitor protein of the enzyme uracil-DNA glycosylase, responsible for uracil excision from DNA. Here, we report that a Staphylococcus pathogenicity island repressor protein called StlSaPIbov1 (Stl) exhibits potent dUTPase inhibition in Mycobacteria. To our knowledge, this is the first indication of a cross-species inhibitor protein for any dUTPase. We demonstrate that the Staphylococcus aureus Stl and the Mycobacterium tuberculosis dUTPase form a stable complex and that in this complex, the enzymatic activity of dUTPase is strongly inhibited. We also found that the expression of the Stl protein in Mycobacterium smegmatis led to highly increased cellular dUTP levels in the mycobacterial cell, this effect being in agreement with its dUTPase inhibitory role. In addition, Stl expression in M. smegmatis drastically decreased colony forming ability, as well, indicating significant perturbation of the phenotype. Therefore, we propose that Stl can be considered to be a cross-species dUTPase inhibitor and may be used as an important reagent in dUTPase inhibition experiments either in vitro/in situ or in vivo.
authors.affiliation: Institute of Enzymology, Research Centre for Natural Sciences (RCNS), Hungarian Academy of Sciences, Budapest, Hungary. Electronic address: hirmondo.rita@ttk.mta.hu.
authors.forename: Rita
authors.initials: R
authors.lastname: Hirmondó
authors.affiliation: Institute of Enzymology, Research Centre for Natural Sciences (RCNS), Hungarian Academy of Sciences, Budapest, Hungary; Department of Applied Biotechnology and Food Sciences, Budapest University of Technology and Economics, Budapest, Hungary.
authors.forename: Judit E
authors.initials: JE
authors.lastname: Szabó
authors.affiliation: Institute of Enzymology, Research Centre for Natural Sciences (RCNS), Hungarian Academy of Sciences, Budapest, Hungary; Department of Applied Biotechnology and Food Sciences, Budapest University of Technology and Economics, Budapest, Hungary.
authors.forename: Kinga
authors.initials: K
authors.lastname: Nyíri
authors.affiliation: Institute of Enzymology, Research Centre for Natural Sciences (RCNS), Hungarian Academy of Sciences, Budapest, Hungary.
authors.forename: Szilvia
authors.initials: S
authors.lastname: Tarjányi
authors.affiliation: Institute of Enzymology, Research Centre for Natural Sciences (RCNS), Hungarian Academy of Sciences, Budapest, Hungary; Department of Applied Biotechnology and Food Sciences, Budapest University of Technology and Economics, Budapest, Hungary.
authors.forename: Paula
authors.initials: P
authors.lastname: Dobrotka
authors.affiliation: Institute of Enzymology, Research Centre for Natural Sciences (RCNS), Hungarian Academy of Sciences, Budapest, Hungary.
authors.forename: Judit
authors.initials: J
authors.lastname: Tóth
authors.affiliation: Institute of Enzymology, Research Centre for Natural Sciences (RCNS), Hungarian Academy of Sciences, Budapest, Hungary; Department of Applied Biotechnology and Food Sciences, Budapest University of Technology and Economics, Budapest, Hungary. Electronic address: vertessy@mail.bme.hu.
authors.forename: Beáta G
authors.initials: BG
authors.lastname: Vértessy
mesh_terms: D001426:Bacterial Proteins / Q000378:metabolism* / Q000494:pharmacology; D004791:Enzyme Inhibitors / Q000378:metabolism* / Q000494:pharmacology; D020102:Mycobacterium smegmatis / Q000201:enzymology*; D009169:Mycobacterium tuberculosis / Q000201:enzymology*; D011755:Pyrophosphatases / Q000037:antagonists & inhibitors*; D013211:Staphylococcus aureus; D014544:Uridine Triphosphate / Q000378:metabolism
chemical_list: D001426:Bacterial Proteins; D004791:Enzyme Inhibitors; D011755:Pyrophosphatases; C020020:dUTP pyrophosphatase; D014544:Uridine Triphosphate
keywords: Genome integrity; High cellular dUTP level; Staphylococcal repressor; dUTPase inhibitor protein