Derepression of SaPIbov1 Is Independent of φNM1 Type 2 dUTPase Activity and Is Inhibited by dUTP and dUMP
| Autor: | Jiri Vlach, Gail E. Christie, Jamil S. Saad, Laura K. Parker, Terje Dokland, Rosanne L.L. Hill |
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| Rok vydání: | 2016 |
| Předmět: |
0301 basic medicine
Staphylococcus aureus Genomic Islands 030106 microbiology Mutant Repressor Virulence medicine.disease_cause Article Bacteriophage 03 medical and health sciences Gene Knockout Techniques Structural Biology medicine Bacteriophages Enzyme Inhibitors Pyrophosphatases Molecular Biology Gene Derepression biology fungi biology.organism_classification Pathogenicity island Molecular biology Repressor Proteins Deoxyuracil Nucleotides Helper Viruses Protein Binding |
| Zdroj: | Journal of molecular biology. 429(10) |
| ISSN: | 1089-8638 |
| Popis: | Staphylococcus aureus is an opportunistic human pathogen able to transfer virulence genes to other cells through the mobilization of S. aureus pathogenicity islands (SaPIs). SaPIs are derepressed and packaged into phage-like transducing particles by helper phages like 80α or φNM1. Phages 80α and φNM1 encode structurally distinct dUTPases, Dut80α (type 1) and DutNM1 (type 2). Both dUTPases can interact with the SaPIbov1 Stl master repressor, leading to derepression and mobilization. That two structurally distinct dUTPases bind the same repressor led us to speculate that dUTPase activity may be important to the derepression process. In type 1 dUTPases, Stl binding is inhibited by dUTP. The purpose of this study was to assess the involvement of dUTP binding and dUTPase activity in derepression by DutNM1. DutNM1 activity mutants were created and tested for dUTPase activity using a novel NMR-based assay. We found that all DutNM1 null activity mutants interacted with the SaPIbov1 Stl C-terminal domain, formed DutNM1–Stl heterodimers, and caused the release of the Pstr promoter. However, promoter release was inhibited in the presence of dUTP or dUMP. We tested two φNM1 mutant phages that had null enzyme activity and found that they could still mobilize SaPIbov1. These results show that only the apo form of DutNM1 is active in Stl derepression and that dUTPase activity is not necessary for the mobilization of SaPIbov1 by DutNM1. |
| Databáze: | OpenAIRE |
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