UV lesions located on the leading strand inhibit DNA replication but do not inhibit SV40 T-antigen helicase activity
Identifieur interne : 000249 ( France/Analysis ); précédent : 000248; suivant : 000250UV lesions located on the leading strand inhibit DNA replication but do not inhibit SV40 T-antigen helicase activity
Auteurs : Xavier Veaute [France] ; Giuseppina Mari-Giglia [France] ; Christopher W. Lawrence [États-Unis] ; Alain Sarasin [France]Source :
- Mutation Research-DNA Repair [ 0921-8777 ] ; 2000.
English descriptors
- Teeft :
- Acad, Adduct, Antigen helicase activity, Assay, Biol, Blockage, Bottom strand, Chem, Dimer, Duplex, Enzyme translocates, Escherichia coli, First component, Fork, Fork progression, Hela, Helicase, Helicase activity, Herpes simplex virus, Human cell, Lesion, Mutation, Mutation research, Natl, Photoproduct, Plasmid, Polymerase, Proc, Progression, Pyrimidine, Reaction products, Replication, Replication assay, Replication fork, Replication forks, Simian, Simian virus, Single lesion, Single photoproduct, Strand, Strand template, Synthetic fork, Template, Unmodified, Unmodified substrate, Unwinding, Unwinding activity, Veaute.
Abstract
Abstract: DNA replication in eucaryotic cells involves a variety of proteins which synthesize the leading and lagging strands in an asymmetric coordinated manner. To analyse the effect of this asymmetry on the translesion synthesis of UV-induced lesions, we have incubated SV40 origin-containing plasmids with a unique site-specific cis,syn-cyclobutane dimer or a pyrimidine-pyrimidone (6-4) photoproduct on either the leading or lagging strand template with DNA replication-competent extracts made from human HeLa cells. Two dimensional agarose gel electrophoresis analyses revealed a strong blockage of fork progression only when the UV lesion is located on the leading strand template. Because DNA helicases are responsible for unwinding duplex DNA ahead of the fork and are then the first component to encounter any potential lesion, we tested the effect of these single photoproducts on the unwinding activity of the SV40 T antigen, the major helicase in our in vitro replication assay. We showed that the activity of the SV40 T-antigen helicase is not inhibited by UV-induced DNA lesions in double-stranded DNA substrate.
Url:
DOI: 10.1016/S0921-8777(99)00052-X
Affiliations:
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ISTEX:B34D211333F5F9478058F8AE131A178CFF0E6D60Le document en format XML
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Lawrence</name><affiliation wicri:level="1"><country xml:lang="fr">États-Unis</country><wicri:regionArea>Department of Biophysics, University of Rochester School of Medicine and Dentistry, Rochester, NY 14642</wicri:regionArea><wicri:noRegion>NY 14642</wicri:noRegion></affiliation></author><author><name sortKey="Sarasin, Alain" sort="Sarasin, Alain" uniqKey="Sarasin A" first="Alain" last="Sarasin">Alain Sarasin</name><affiliation wicri:level="3"><country xml:lang="fr">France</country><wicri:regionArea>Institut de Recherches sur le Cancer, UPR 42 CNRS, Laboratoire de Génétique Moléculaire, BP 8, 7, rue Guy-Moquet, 94801 Villejuif Cedex</wicri:regionArea><placeName><region type="region" nuts="2">Île-de-France</region><settlement type="city">Villejuif</settlement></placeName></affiliation></author></analytic><monogr></monogr><series><title level="j">Mutation Research-DNA Repair</title><title level="j" type="abbrev">MUTDNA</title><idno type="ISSN">0921-8777</idno><imprint><publisher>ELSEVIER</publisher><date type="published" when="2000">2000</date><biblScope unit="volume">459</biblScope><biblScope unit="issue">1</biblScope><biblScope unit="page" from="19">19</biblScope><biblScope unit="page" to="28">28</biblScope></imprint><idno type="ISSN">0921-8777</idno></series></biblStruct></sourceDesc><seriesStmt><idno type="ISSN">0921-8777</idno></seriesStmt></fileDesc><profileDesc><textClass><keywords scheme="Teeft" xml:lang="en"><term>Acad</term><term>Adduct</term><term>Antigen helicase activity</term><term>Assay</term><term>Biol</term><term>Blockage</term><term>Bottom strand</term><term>Chem</term><term>Dimer</term><term>Duplex</term><term>Enzyme translocates</term><term>Escherichia coli</term><term>First component</term><term>Fork</term><term>Fork progression</term><term>Hela</term><term>Helicase</term><term>Helicase activity</term><term>Herpes simplex virus</term><term>Human cell</term><term>Lesion</term><term>Mutation</term><term>Mutation research</term><term>Natl</term><term>Photoproduct</term><term>Plasmid</term><term>Polymerase</term><term>Proc</term><term>Progression</term><term>Pyrimidine</term><term>Reaction products</term><term>Replication</term><term>Replication assay</term><term>Replication fork</term><term>Replication forks</term><term>Simian</term><term>Simian virus</term><term>Single lesion</term><term>Single photoproduct</term><term>Strand</term><term>Strand template</term><term>Synthetic fork</term><term>Template</term><term>Unmodified</term><term>Unmodified substrate</term><term>Unwinding</term><term>Unwinding activity</term><term>Veaute</term></keywords></textClass><langUsage><language ident="en">en</language></langUsage></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">Abstract: DNA replication in eucaryotic cells involves a variety of proteins which synthesize the leading and lagging strands in an asymmetric coordinated manner. To analyse the effect of this asymmetry on the translesion synthesis of UV-induced lesions, we have incubated SV40 origin-containing plasmids with a unique site-specific cis,syn-cyclobutane dimer or a pyrimidine-pyrimidone (6-4) photoproduct on either the leading or lagging strand template with DNA replication-competent extracts made from human HeLa cells. Two dimensional agarose gel electrophoresis analyses revealed a strong blockage of fork progression only when the UV lesion is located on the leading strand template. Because DNA helicases are responsible for unwinding duplex DNA ahead of the fork and are then the first component to encounter any potential lesion, we tested the effect of these single photoproducts on the unwinding activity of the SV40 T antigen, the major helicase in our in vitro replication assay. We showed that the activity of the SV40 T-antigen helicase is not inhibited by UV-induced DNA lesions in double-stranded DNA substrate.</div></front></TEI><affiliations><list><country><li>France</li><li>États-Unis</li></country><region><li>Île-de-France</li></region><settlement><li>Villejuif</li></settlement></list><tree><country name="France"><region name="Île-de-France"><name sortKey="Veaute, Xavier" sort="Veaute, Xavier" uniqKey="Veaute X" first="Xavier" last="Veaute">Xavier Veaute</name></region><name sortKey="Mari Giglia, Giuseppina" sort="Mari Giglia, Giuseppina" uniqKey="Mari Giglia G" first="Giuseppina" last="Mari-Giglia">Giuseppina Mari-Giglia</name><name sortKey="Sarasin, Alain" sort="Sarasin, Alain" uniqKey="Sarasin A" first="Alain" last="Sarasin">Alain Sarasin</name></country><country name="États-Unis"><noRegion><name sortKey="Lawrence, Christopher W" sort="Lawrence, Christopher W" uniqKey="Lawrence C" first="Christopher W" last="Lawrence">Christopher W. 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