Functional Characterization of Uracil-DNA Glycosylase from Staphylococcus aureus
Abstract
Background/Objectives: Uracil-DNA glycosylase (UNG) initiates base excision repair by removing uracil from DNA. Although Family 1 UNGs are well characterized in model organisms, the biochemical and functional properties of Staphylococcus aureus UNG (SaUNG) is not well characterized. This study aimed to characterize SaUNG. Methods: Recombinant wild-type and mutant SaUNG proteins were examined using biochemical and kinetic assays, site-directed mutagenesis, genetic complementation in Escherichia coli, and in vitro reconstitution of S. aureus base excision repair. Results: SaUNG exhibited maximal activity at pH 8.0–8.5, 37–45 °C, and 100–150 mM KCl and lacked AP-lyase activity. Catalytic efficiencies (kcat/Km) were 1.39 × 109, 3.79 × 108, and 2.93 × 108 M−1·min−1 for ssU, U:G, and U:A, respectively, establishing the preference ssU > U:G > U:A. H180A retained residual activity, whereas simultaneous substitution of Asp59 and His180 abolished detectable activity. Complete inhibition confirmed conserved biochemical properties of Family 1 UNGs. SaUNG complemented the mutation avoidance defect of an E. coli (ung− mug−) mutant and initiated uracil repair in a reconstituted S. aureus base excision repair system. Conclusions: SaUNG is a canonical Family 1 UNG with experimentally characterized biochemical properties, catalytic determinants, and biological function in uracil repair.
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BibTeX
@article{koreneva2026functional,
title = {{Functional Characterization of Uracil-DNA Glycosylase from Staphylococcus aureus}},
author = {Arina Koreneva and Albina Sugirova and Bakhtiyar Yakupov and Maria V Safonkina and Anton V Endutkin and Dmitry O Zharkov and Murat Saparbaev and Sailau Abeldenov and Aigerim Turgimbayeva},
journal = {Pathogens},
year = {2026},
month = sep,
doi = {10.3390/pathogens15101012},
url = {https://hal.science/hal-05772353v1},
note = {CC BY 4.0},
}
RIS
TY - JOUR
TI - Functional Characterization of Uracil-DNA Glycosylase from Staphylococcus aureus
AU - Arina Koreneva
AU - Albina Sugirova
AU - Bakhtiyar Yakupov
AU - Maria V Safonkina
AU - Anton V Endutkin
AU - Dmitry O Zharkov
AU - Murat Saparbaev
AU - Sailau Abeldenov
AU - Aigerim Turgimbayeva
PY - 2026
DA - 2026/09/24
JO - Pathogens
DO - 10.3390/pathogens15101012
UR - https://hal.science/hal-05772353v1
AB - Background/Objectives: Uracil-DNA glycosylase (UNG) initiates base excision repair by removing uracil from DNA. Although Family 1 UNGs are well characterized in model organisms, the biochemical and functional properties of Staphylococcus aureus UNG (SaUNG) is not well characterized. This study aimed to characterize SaUNG. Methods: Recombinant wild-type and mutant SaUNG proteins were examined using biochemical and kinetic assays, site-directed mutagenesis, genetic complementation in Escherichia coli, and in vitro reconstitution of S. aureus base excision repair. Results: SaUNG exhibited maximal activity at pH 8.0–8.5, 37–45 °C, and 100–150 mM KCl and lacked AP-lyase activity. Catalytic efficiencies (kcat/Km) were 1.39 × 109, 3.79 × 108, and 2.93 × 108 M−1·min−1 for ssU, U:G, and U:A, respectively, establishing the preference ssU > U:G > U:A. H180A retained residual activity, whereas simultaneous substitution of Asp59 and His180 abolished detectable activity. Complete inhibition confirmed conserved biochemical properties of Family 1 UNGs. SaUNG complemented the mutation avoidance defect of an E. coli (ung− mug−) mutant and initiated uracil repair in a reconstituted S. aureus base excision repair system. Conclusions: SaUNG is a canonical Family 1 UNG with experimentally characterized biochemical properties, catalytic determinants, and biological function in uracil repair.
SN - 2076-0817
ER -