Enzymatic analysis of oligonucleotides containing cyclobutane pyrimidine photodimers with a cleaved intradimer phosphodiester linkage.
Identifieur interne : 000216 ( Ncbi/Checkpoint ); précédent : 000215; suivant : 000217Enzymatic analysis of oligonucleotides containing cyclobutane pyrimidine photodimers with a cleaved intradimer phosphodiester linkage.
Auteurs : M. Liuzzi [Canada] ; M C PatersonSource :
- The Journal of biological chemistry [ 0021-9258 ] ; 1992.
Descripteurs français
- KwdFr :
- MESH :
- analyse : Dimères de pyrimidine.
- enzymologie : Escherichia coli.
- métabolisme : DNA nucleotidylexotransferase, Deoxyribodipyrimidine photo-lyase, Phosphodiesterases.
- Altération de l'ADN, Phosphates, Réparation de l'ADN, Techniques in vitro.
English descriptors
- KwdEn :
- MESH :
- chemical , analysis : Pyrimidine Dimers.
- chemical , metabolism : DNA Nucleotidylexotransferase, Deoxyribodipyrimidine Photo-Lyase, Phosphoric Diester Hydrolases.
- enzymology : Escherichia coli.
- DNA Damage, DNA Repair, In Vitro Techniques, Phosphates.
Abstract
Our recent studies indicate that enzymatic hydrolysis of the intradimer phosphodiester linkage constitutes an early reaction in processing UV light-induced cis-syn-cyclobutane pyrimidine dimers in cultured human fibroblasts. Before characterizing the resultant modified dimer sites in cellular DNA, it is necessary to establish experimental conditions that can distinguish backbone-nicked from intact dimers. We thus constructed a model substrate, i.e. p(dT) 10 <> p(dT)10 containing a dimer with a ruptured sugar-phosphate bond, and determined the products of its reaction with snake venom phosphodiesterase and alkaline phosphatase, an enzymatic digestion mixture known to release dimers from UV-treated poly(dA).poly(dT) within trinucleotides with the photoproduct intact at the 3'-end (d-TpT
T). The model substrate was prepared by (i) end labeling p(dT)9 using terminal deoxynucleotidyltransferase and [3H]thymine-labeled TTP; and (ii) annealing the chromatographically purified p(dT)10 oligomers to poly(dA) followed by UV (290 nm)-induced ligation. Photoligated 20-mers with one radioactive and modified internal dimer were isolated and enzymatically digested. High performance liquid chromatographic analysis of the reaction products revealed a novel trithymidylate with its backbone severed at the 3'-terminus (d-TpT<>dT), demonstrating that this procedure could discriminate between intact and modified dimers. The procedure was then exploited to show that (i) Escherichia coli DNA photolyase can monomerize, albeit inefficiently, backbone-ruptured dimers; and (ii) phage T4 polynucleotide kinase can catalyze the phosphorylation of d-TpT<>dT, thus facilitating the development of a sensitive postlabeling assay suitable for modified dimer detection under biologically relevant conditions.
PubMed: 1331055
Affiliations:
Links toward previous steps (curation, corpus...)
- to stream PubMed, to step Corpus: 002942
- to stream PubMed, to step Curation: 002942
- to stream PubMed, to step Checkpoint: 002812
- to stream Ncbi, to step Merge: 000216
- to stream Ncbi, to step Curation: 000216
Links to Exploration step
pubmed:1331055Le document en format XML
<record><TEI><teiHeader><fileDesc><titleStmt><title xml:lang="en">Enzymatic analysis of oligonucleotides containing cyclobutane pyrimidine photodimers with a cleaved intradimer phosphodiester linkage.</title><author><name sortKey="Liuzzi, M" sort="Liuzzi, M" uniqKey="Liuzzi M" first="M" last="Liuzzi">M. Liuzzi</name><affiliation wicri:level="1"><nlm:affiliation>Department of Medicine, Cross Cancer Institute, Edmonton, Alberta, Canada.</nlm:affiliation><country xml:lang="fr">Canada</country><wicri:regionArea>Department of Medicine, Cross Cancer Institute, Edmonton, Alberta</wicri:regionArea><wicri:noRegion>Alberta</wicri:noRegion></affiliation></author><author><name sortKey="Paterson, M C" sort="Paterson, M C" uniqKey="Paterson M" first="M C" last="Paterson">M C Paterson</name></author></titleStmt><publicationStmt><idno type="wicri:source">PubMed</idno><date when="1992">1992</date><idno type="RBID">pubmed:1331055</idno><idno type="pmid">1331055</idno><idno type="wicri:Area/PubMed/Corpus">002942</idno><idno type="wicri:explorRef" wicri:stream="PubMed" wicri:step="Corpus" wicri:corpus="PubMed">002942</idno><idno type="wicri:Area/PubMed/Curation">002942</idno><idno type="wicri:explorRef" wicri:stream="PubMed" wicri:step="Curation">002942</idno><idno type="wicri:Area/PubMed/Checkpoint">002812</idno><idno type="wicri:explorRef" wicri:stream="Checkpoint" wicri:step="PubMed">002812</idno><idno type="wicri:Area/Ncbi/Merge">000216</idno><idno type="wicri:Area/Ncbi/Curation">000216</idno><idno type="wicri:Area/Ncbi/Checkpoint">000216</idno></publicationStmt><sourceDesc><biblStruct><analytic><title xml:lang="en">Enzymatic analysis of oligonucleotides containing cyclobutane pyrimidine photodimers with a cleaved intradimer phosphodiester linkage.</title><author><name sortKey="Liuzzi, M" sort="Liuzzi, M" uniqKey="Liuzzi M" first="M" last="Liuzzi">M. Liuzzi</name><affiliation wicri:level="1"><nlm:affiliation>Department of Medicine, Cross Cancer Institute, Edmonton, Alberta, Canada.</nlm:affiliation><country xml:lang="fr">Canada</country><wicri:regionArea>Department of Medicine, Cross Cancer Institute, Edmonton, Alberta</wicri:regionArea><wicri:noRegion>Alberta</wicri:noRegion></affiliation></author><author><name sortKey="Paterson, M C" sort="Paterson, M C" uniqKey="Paterson M" first="M C" last="Paterson">M C Paterson</name></author></analytic><series><title level="j">The Journal of biological chemistry</title><idno type="ISSN">0021-9258</idno><imprint><date when="1992" type="published">1992</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>DNA Damage</term><term>DNA Nucleotidylexotransferase (metabolism)</term><term>DNA Repair</term><term>Deoxyribodipyrimidine Photo-Lyase (metabolism)</term><term>Escherichia coli (enzymology)</term><term>In Vitro Techniques</term><term>Phosphates</term><term>Phosphoric Diester Hydrolases (metabolism)</term><term>Pyrimidine Dimers (analysis)</term></keywords><keywords scheme="KwdFr" xml:lang="fr"><term>Altération de l'ADN</term><term>DNA nucleotidylexotransferase (métabolisme)</term><term>Deoxyribodipyrimidine photo-lyase (métabolisme)</term><term>Dimères de pyrimidine (analyse)</term><term>Escherichia coli (enzymologie)</term><term>Phosphates</term><term>Phosphodiesterases (métabolisme)</term><term>Réparation de l'ADN</term><term>Techniques in vitro</term></keywords><keywords scheme="MESH" type="chemical" qualifier="analysis" xml:lang="en"><term>Pyrimidine Dimers</term></keywords><keywords scheme="MESH" type="chemical" qualifier="metabolism" xml:lang="en"><term>DNA Nucleotidylexotransferase</term><term>Deoxyribodipyrimidine Photo-Lyase</term><term>Phosphoric Diester Hydrolases</term></keywords><keywords scheme="MESH" qualifier="analyse" xml:lang="fr"><term>Dimères de pyrimidine</term></keywords><keywords scheme="MESH" qualifier="enzymologie" xml:lang="fr"><term>Escherichia coli</term></keywords><keywords scheme="MESH" qualifier="enzymology" xml:lang="en"><term>Escherichia coli</term></keywords><keywords scheme="MESH" qualifier="métabolisme" xml:lang="fr"><term>DNA nucleotidylexotransferase</term><term>Deoxyribodipyrimidine photo-lyase</term><term>Phosphodiesterases</term></keywords><keywords scheme="MESH" xml:lang="en"><term>DNA Damage</term><term>DNA Repair</term><term>In Vitro Techniques</term><term>Phosphates</term></keywords><keywords scheme="MESH" xml:lang="fr"><term>Altération de l'ADN</term><term>Phosphates</term><term>Réparation de l'ADN</term><term>Techniques in vitro</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">Our recent studies indicate that enzymatic hydrolysis of the intradimer phosphodiester linkage constitutes an early reaction in processing UV light-induced cis-syn-cyclobutane pyrimidine dimers in cultured human fibroblasts. Before characterizing the resultant modified dimer sites in cellular DNA, it is necessary to establish experimental conditions that can distinguish backbone-nicked from intact dimers. We thus constructed a model substrate, i.e. p(dT) 10 <> p(dT)10 containing a dimer with a ruptured sugar-phosphate bond, and determined the products of its reaction with snake venom phosphodiesterase and alkaline phosphatase, an enzymatic digestion mixture known to release dimers from UV-treated poly(dA).poly(dT) within trinucleotides with the photoproduct intact at the 3'-end (d-TpTT). The model substrate was prepared by (i) end labeling p(dT)9 using terminal deoxynucleotidyltransferase and [3H]thymine-labeled TTP; and (ii) annealing the chromatographically purified p(dT)10 oligomers to poly(dA) followed by UV (290 nm)-induced ligation. Photoligated 20-mers with one radioactive and modified internal dimer were isolated and enzymatically digested. High performance liquid chromatographic analysis of the reaction products revealed a novel trithymidylate with its backbone severed at the 3'-terminus (d-TpT<>dT), demonstrating that this procedure could discriminate between intact and modified dimers. The procedure was then exploited to show that (i) Escherichia coli DNA photolyase can monomerize, albeit inefficiently, backbone-ruptured dimers; and (ii) phage T4 polynucleotide kinase can catalyze the phosphorylation of d-TpT<>dT, thus facilitating the development of a sensitive postlabeling assay suitable for modified dimer detection under biologically relevant conditions.</div>
</front></TEI><affiliations><list><country><li>Canada</li></country></list><tree><noCountry><name sortKey="Paterson, M C" sort="Paterson, M C" uniqKey="Paterson M" first="M C" last="Paterson">M C Paterson</name></noCountry><country name="Canada"><noRegion><name sortKey="Liuzzi, M" sort="Liuzzi, M" uniqKey="Liuzzi M" first="M" last="Liuzzi">M. Liuzzi</name></noRegion></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
EXPLOR_STEP=$WICRI_ROOT/Sante/explor/MersV1/Data/Ncbi/Checkpoint
HfdSelect -h $EXPLOR_STEP/biblio.hfd -nk 000216 | SxmlIndent | more
Ou
HfdSelect -h $EXPLOR_AREA/Data/Ncbi/Checkpoint/biblio.hfd -nk 000216 | SxmlIndent | more
Pour mettre un lien sur cette page dans le réseau Wicri
{{Explor lien
|wiki= Sante
|area= MersV1
|flux= Ncbi
|étape= Checkpoint
|type= RBID
|clé= pubmed:1331055
|texte= Enzymatic analysis of oligonucleotides containing cyclobutane pyrimidine photodimers with a cleaved intradimer phosphodiester linkage.
}}
Pour générer des pages wiki
HfdIndexSelect -h $EXPLOR_AREA/Data/Ncbi/Checkpoint/RBID.i -Sk "pubmed:1331055" \
| HfdSelect -Kh $EXPLOR_AREA/Data/Ncbi/Checkpoint/biblio.hfd \
| NlmPubMed2Wicri -a MersV1
| This area was generated with Dilib version V0.6.33. |  |