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7,8-dihydro-8-oxoadenine, a highly mutagenic adduct, is repaired by Escherichia coli and human mismatch-specific uracil/thymine-DNA glycosylases

Identifieur interne : 000773 ( Main/Exploration ); précédent : 000772; suivant : 000774

7,8-dihydro-8-oxoadenine, a highly mutagenic adduct, is repaired by Escherichia coli and human mismatch-specific uracil/thymine-DNA glycosylases

Auteurs : Ibtissam Talhaoui [France] ; Sophie Couvé [France] ; Alexander A. Ishchenko [France] ; Christophe Kunz ; Primo Sch R ; Murat Saparbaev [France]

Source :

RBID : PMC:3553953

Abstract

Hydroxyl radicals predominantly react with the C8 of purines forming 7,8-dihydro-8-oxoguanine (8oxoG) and 7,8-dihydro-8-oxoadenine (8oxoA) adducts, which are highly mutagenic in mammalian cells. The majority of oxidized DNA bases are removed by DNA glycosylases in the base excision repair pathway. Here, we report for the first time that human thymine-DNA glycosylase (hTDG) and Escherichia coli mismatch-specific uracil-DNA glycosylase (MUG) can remove 8oxoA from 8oxoA•T, 8oxoA•G and 8oxoA•C pairs. Comparison of the kinetic parameters of the reaction indicates that full-length hTDG excises 8oxoA, 3,N4-ethenocytosine (εC) and T with similar efficiency (kmax = 0.35, 0.36 and 0.16 min−1, respectively) and is more proficient as compared with its bacterial homologue MUG. The N-terminal domain of the hTDG protein is essential for 8oxoA–DNA glycosylase activity, but not for εC repair. Interestingly, the TDG status had little or no effect on the proliferation rate of mouse embryonic fibroblasts after exposure to γ-irradiation. Nevertheless, using whole cell-free extracts from the DNA glycosylase-deficient murine embryonic fibroblasts and E. coli, we demonstrate that the excision of 8oxoA from 8oxoA•T and 8oxoA•G has an absolute requirement for TDG and MUG, respectively. The data establish that MUG and TDG can counteract the genotoxic effects of 8oxoA residues in vivo.


Url:
DOI: 10.1093/nar/gks1149
PubMed: 23209024
PubMed Central: 3553953


Affiliations:


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Le document en format XML

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<p>Hydroxyl radicals predominantly react with the C
<sub>8</sub>
of purines forming 7,8-dihydro-8-oxoguanine (8oxoG) and 7,8-dihydro-8-oxoadenine (8oxoA) adducts, which are highly mutagenic in mammalian cells. The majority of oxidized DNA bases are removed by DNA glycosylases in the base excision repair pathway. Here, we report for the first time that human thymine-DNA glycosylase (hTDG) and
<italic>Escherichia coli</italic>
mismatch-specific uracil-DNA glycosylase (MUG) can remove 8oxoA from 8oxoA•T, 8oxoA•G and 8oxoA•C pairs. Comparison of the kinetic parameters of the reaction indicates that full-length hTDG excises 8oxoA, 3,
<italic>N</italic>
<sup>4</sup>
-ethenocytosine (εC) and T with similar efficiency (
<italic>k</italic>
<sub>max</sub>
= 0.35, 0.36 and 0.16 min
<sup>−1</sup>
, respectively) and is more proficient as compared with its bacterial homologue MUG. The N-terminal domain of the hTDG protein is essential for 8oxoA–DNA glycosylase activity, but not for εC repair. Interestingly, the TDG status had little or no effect on the proliferation rate of mouse embryonic fibroblasts after exposure to γ-irradiation. Nevertheless, using whole cell-free extracts from the DNA glycosylase-deficient murine embryonic fibroblasts and
<italic>E. coli</italic>
, we demonstrate that the excision of 8oxoA from 8oxoA•T and 8oxoA•G has an absolute requirement for TDG and MUG, respectively. The data establish that MUG and TDG can counteract the genotoxic effects of 8oxoA residues
<italic>in vivo</italic>
.</p>
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<name sortKey="Otterlei, M" uniqKey="Otterlei M">M Otterlei</name>
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<author>
<name sortKey="Nilsen, H" uniqKey="Nilsen H">H Nilsen</name>
</author>
<author>
<name sortKey="Skorpen, F" uniqKey="Skorpen F">F Skorpen</name>
</author>
<author>
<name sortKey="Aas, Pa" uniqKey="Aas P">PA Aas</name>
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<author>
<name sortKey="Hagen, L" uniqKey="Hagen L">L Hagen</name>
</author>
<author>
<name sortKey="Krokan, He" uniqKey="Krokan H">HE Krokan</name>
</author>
<author>
<name sortKey="Slupphaug, G" uniqKey="Slupphaug G">G Slupphaug</name>
</author>
</analytic>
</biblStruct>
<biblStruct>
<analytic>
<author>
<name sortKey="Vasovcak, P" uniqKey="Vasovcak P">P Vasovcak</name>
</author>
<author>
<name sortKey="Krepelova, A" uniqKey="Krepelova A">A Krepelova</name>
</author>
<author>
<name sortKey="Menigatti, M" uniqKey="Menigatti M">M Menigatti</name>
</author>
<author>
<name sortKey="Puchmajerova, A" uniqKey="Puchmajerova A">A Puchmajerova</name>
</author>
<author>
<name sortKey="Skapa, P" uniqKey="Skapa P">P Skapa</name>
</author>
<author>
<name sortKey="Augustinakova, A" uniqKey="Augustinakova A">A Augustinakova</name>
</author>
<author>
<name sortKey="Amann, G" uniqKey="Amann G">G Amann</name>
</author>
<author>
<name sortKey="Wernstedt, A" uniqKey="Wernstedt A">A Wernstedt</name>
</author>
<author>
<name sortKey="Jiricny, J" uniqKey="Jiricny J">J Jiricny</name>
</author>
<author>
<name sortKey="Marra, G" uniqKey="Marra G">G Marra</name>
</author>
</analytic>
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</TEI>
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<list>
<country>
<li>France</li>
</country>
<region>
<li>Île-de-France</li>
</region>
<settlement>
<li>Villejuif</li>
</settlement>
<orgName>
<li>Université Paris-Sud</li>
</orgName>
</list>
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<name sortKey="Kunz, Christophe" sort="Kunz, Christophe" uniqKey="Kunz C" first="Christophe" last="Kunz">Christophe Kunz</name>
<name sortKey="Sch R, Primo" sort="Sch R, Primo" uniqKey="Sch R P" first="Primo" last="Sch R">Primo Sch R</name>
</noCountry>
<country name="France">
<region name="Île-de-France">
<name sortKey="Talhaoui, Ibtissam" sort="Talhaoui, Ibtissam" uniqKey="Talhaoui I" first="Ibtissam" last="Talhaoui">Ibtissam Talhaoui</name>
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<name sortKey="Couve, Sophie" sort="Couve, Sophie" uniqKey="Couve S" first="Sophie" last="Couvé">Sophie Couvé</name>
<name sortKey="Couve, Sophie" sort="Couve, Sophie" uniqKey="Couve S" first="Sophie" last="Couvé">Sophie Couvé</name>
<name sortKey="Ishchenko, Alexander A" sort="Ishchenko, Alexander A" uniqKey="Ishchenko A" first="Alexander A." last="Ishchenko">Alexander A. Ishchenko</name>
<name sortKey="Saparbaev, Murat" sort="Saparbaev, Murat" uniqKey="Saparbaev M" first="Murat" last="Saparbaev">Murat Saparbaev</name>
</country>
</tree>
</affiliations>
</record>

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