Consensus sequences for good and poor removal of uracil from double stranded DNA by uracil-DNA glycosylase
Identifieur interne : 004587 ( Main/Exploration ); précédent : 004586; suivant : 004588Consensus sequences for good and poor removal of uracil from double stranded DNA by uracil-DNA glycosylase
Auteurs : Ingrid Eftedal [Norvège] ; Per H. Guddal ; Geir Slupphaug [Norvège] ; Gunnar Volden [Norvège] ; Hans E. Krokan [Norvège]Source :
- Nucleic Acids Research [ 0305-1048 ] ; 1993.
Abstract
We have purified uracil DNA-glycosylase (UDG) from calf thymus 32,OOO-fold and studied Its biochemical properties, including sequence specificity. The enzyme is apparently closely related to human UDG, since it was recognised by a polyclonal antibody directed towards human UDG. SDS-PAGE and western analysis Indicate an apparent Mr = 27,500. Bovine UDG has a 1.7-fold preference for single stranded over double stranded DNA as a substrate. Sequence specificity for uracil removal from dsDNA was examined for bovine and Escherichla coll UDG, using DNA containing less than one dUMP residper 100 nucleotides and synthetic oligonucleotides containing one dUMP residue. Comparative studies Involving about 40 uracil sites Indicated similar specificities for both UDGs. We found more than a 10-fold difference In rates of uracil removal between different sequences. 5’-G/cUT-3’ and 5’-G/cUG/c-3’ were consensus sequences for poor repair whereas 5’-A/TUAA/T-3’ was a consensus for good repair. Sequence specificity was verified in double stranded oligonucleotides, but not in single stranded ones, suggesting that the structure of the double stranded DNA helix has Influence on sequence specificity. Rate of uracil removal appeared to be slightly faster from U:A base pairs as compared to U:G mis-matches. The results indicate that sequence specific repair may be a determinant to be considered in mutagenesis.
Url:
DOI: 10.1093/nar/21.9.2095
Affiliations:
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Guddal</name><affiliation><wicri:noCountry code="subField">Tromsø</wicri:noCountry></affiliation></author><author><name sortKey="Slupphaug, Geir" sort="Slupphaug, Geir" uniqKey="Slupphaug G" first="Geir" last="Slupphaug">Geir Slupphaug</name><affiliation wicri:level="1"><country>Norvège</country><placeName><settlement type="city">Trondheim</settlement><region type="région" nuts="2">Trøndelag</region></placeName><wicri:regionArea>UNIGEN Center for Molecular Biology</wicri:regionArea></affiliation></author><author><name sortKey="Volden, Gunnar" sort="Volden, Gunnar" uniqKey="Volden G" first="Gunnar" last="Volden">Gunnar Volden</name><affiliation wicri:level="1"><country xml:lang="fr">Norvège</country><wicri:regionArea>Department of Dermatology, University of Trondheim N-7006 Trondheim</wicri:regionArea><wicri:noRegion>University of Trondheim N-7006 Trondheim</wicri:noRegion></affiliation></author><author><name sortKey="Krokan, Hans E" sort="Krokan, Hans E" uniqKey="Krokan H" first="Hans E." last="Krokan">Hans E. 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The enzyme is apparently closely related to human UDG, since it was recognised by a polyclonal antibody directed towards human UDG. SDS-PAGE and western analysis Indicate an apparent Mr = 27,500. Bovine UDG has a 1.7-fold preference for single stranded over double stranded DNA as a substrate. Sequence specificity for uracil removal from dsDNA was examined for bovine and Escherichla coll UDG, using DNA containing less than one dUMP residper 100 nucleotides and synthetic oligonucleotides containing one dUMP residue. Comparative studies Involving about 40 uracil sites Indicated similar specificities for both UDGs. We found more than a 10-fold difference In rates of uracil removal between different sequences. 5’-G/cUT-3’ and 5’-G/cUG/c-3’ were consensus sequences for poor repair whereas 5’-A/TUAA/T-3’ was a consensus for good repair. Sequence specificity was verified in double stranded oligonucleotides, but not in single stranded ones, suggesting that the structure of the double stranded DNA helix has Influence on sequence specificity. Rate of uracil removal appeared to be slightly faster from U:A base pairs as compared to U:G mis-matches. The results indicate that sequence specific repair may be a determinant to be considered in mutagenesis.</div></front></TEI><affiliations><list><country><li>Norvège</li></country><region><li>Trøndelag</li></region><settlement><li>Trondheim</li></settlement></list><tree><noCountry><name sortKey="Guddal, Per H" sort="Guddal, Per H" uniqKey="Guddal P" first="Per H." last="Guddal">Per H. Guddal</name></noCountry><country name="Norvège"><region name="Trøndelag"><name sortKey="Eftedal, Ingrid" sort="Eftedal, Ingrid" uniqKey="Eftedal I" first="Ingrid" last="Eftedal">Ingrid Eftedal</name></region><name sortKey="Krokan, Hans E" sort="Krokan, Hans E" uniqKey="Krokan H" first="Hans E." last="Krokan">Hans E. Krokan</name><name sortKey="Slupphaug, Geir" sort="Slupphaug, Geir" uniqKey="Slupphaug G" first="Geir" last="Slupphaug">Geir Slupphaug</name><name sortKey="Volden, Gunnar" sort="Volden, Gunnar" uniqKey="Volden G" first="Gunnar" last="Volden">Gunnar Volden</name></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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