Research output: Contribution to journal › Article › peer-review
DNA glycosylases for 8-oxoguanine repair in Staphylococcus aureus. / Endutkin, Anton V.; Panferova, Elena P.; Barmatov, Alexander E. et al.
In: DNA Repair, Vol. 105, 103160, 09.2021, p. 103160.Research output: Contribution to journal › Article › peer-review
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TY - JOUR
T1 - DNA glycosylases for 8-oxoguanine repair in Staphylococcus aureus
AU - Endutkin, Anton V.
AU - Panferova, Elena P.
AU - Barmatov, Alexander E.
AU - Zharkov, Dmitry O.
N1 - Funding Information: This research was supported by Russian Science Foundation (grant 19-74-00068 to A.V.E.). Partial salary support from the Russian Ministry of Science and Higher Education (State funded budget projects АААА-А17-117020210023-1 and FSUS-2020-0035) is acknowledged. DNA sequencing was performed at the SB RAS Genomics Core Facility. Publisher Copyright: © 2021 Elsevier B.V.
PY - 2021/9
Y1 - 2021/9
N2 - GO system is part of base excision DNA repair and is required for the correct repair of 8-oxoguanine (8-oxoG), one of the most abundant oxidative lesions. Due to the ability of 8-oxoG to mispair with A, this base is highly mutagenic, and its repair requires two enzymes: Fpg that removes 8-oxoG from 8-oxoG:C pairs, and MutY that excises the normal A from 8-oxoG:A mispairs. Here we characterize the properties of putative GO system DNA glycosylases from Staphylococcus aureus, an important human opportunistic pathogen that causes hospital infections and presents a serious health concern due to quick spread of antibiotic-resistant strains. In addition to Fpg and MutY from the reference NCTC 8325 strain (SauFpg1 and SauMutY), we have also studied an Fpg homolog from a multidrug-resistant C0673 isolate (SauFpg2), which is different from SauFpg1 in its sequence. Both SauFpg enzymes showed the highest activity at pH 7.0–9.0 and NaCl concentrations 25–75 mM (SauFpg1) or 50–100 mM (SauFpg2), whereas SauMutY was active at a broad pH range and had a salt optimum at ∼75 mM NaCl. Both SauFpg1 and SauFpg2 bound and cleaved duplexes containing 8-oxoG, 5-hydroxyuracil, 5,6-dihydrouracil or apurinic/apyrimidinic site paired with C, T, or G, but not with A. For SauFpg1 and SauFpg2, 8-oxoG was the best substrate tested, and 5,6-dihydrouracil was the worst one. SauMutY efficiently excised adenine from duplex substrates containing A:8-oxoG or A:G pairs. SauFpg enzymes were readily trapped on DNA by NaBH4 treatment, indicating formation of a Schiff base reaction intermediate. Surprisingly, SauMutY was also trapped significantly better than its E. coli homolog. All three S. aureus GO glycosylases drastically reduced spontaneous mutagenesis when expressed in an fpg mutY E. coli double mutant. Overall, we conclude that S. aureus possesses an active GO system, which could possibly be targeted for sensitization of this pathogen to oxidative stress.
AB - GO system is part of base excision DNA repair and is required for the correct repair of 8-oxoguanine (8-oxoG), one of the most abundant oxidative lesions. Due to the ability of 8-oxoG to mispair with A, this base is highly mutagenic, and its repair requires two enzymes: Fpg that removes 8-oxoG from 8-oxoG:C pairs, and MutY that excises the normal A from 8-oxoG:A mispairs. Here we characterize the properties of putative GO system DNA glycosylases from Staphylococcus aureus, an important human opportunistic pathogen that causes hospital infections and presents a serious health concern due to quick spread of antibiotic-resistant strains. In addition to Fpg and MutY from the reference NCTC 8325 strain (SauFpg1 and SauMutY), we have also studied an Fpg homolog from a multidrug-resistant C0673 isolate (SauFpg2), which is different from SauFpg1 in its sequence. Both SauFpg enzymes showed the highest activity at pH 7.0–9.0 and NaCl concentrations 25–75 mM (SauFpg1) or 50–100 mM (SauFpg2), whereas SauMutY was active at a broad pH range and had a salt optimum at ∼75 mM NaCl. Both SauFpg1 and SauFpg2 bound and cleaved duplexes containing 8-oxoG, 5-hydroxyuracil, 5,6-dihydrouracil or apurinic/apyrimidinic site paired with C, T, or G, but not with A. For SauFpg1 and SauFpg2, 8-oxoG was the best substrate tested, and 5,6-dihydrouracil was the worst one. SauMutY efficiently excised adenine from duplex substrates containing A:8-oxoG or A:G pairs. SauFpg enzymes were readily trapped on DNA by NaBH4 treatment, indicating formation of a Schiff base reaction intermediate. Surprisingly, SauMutY was also trapped significantly better than its E. coli homolog. All three S. aureus GO glycosylases drastically reduced spontaneous mutagenesis when expressed in an fpg mutY E. coli double mutant. Overall, we conclude that S. aureus possesses an active GO system, which could possibly be targeted for sensitization of this pathogen to oxidative stress.
KW - 8-Oxoguanine
KW - DNA damage
KW - DNA glycosylases
KW - DNA repair
KW - GO system
KW - Staphylococcus aureus
UR - http://www.scopus.com/inward/record.url?scp=85111071193&partnerID=8YFLogxK
U2 - 10.1016/j.dnarep.2021.103160
DO - 10.1016/j.dnarep.2021.103160
M3 - Article
C2 - 34192601
AN - SCOPUS:85111071193
VL - 105
SP - 103160
JO - DNA Repair
JF - DNA Repair
SN - 1568-7864
M1 - 103160
ER -
ID: 29131285