The Mammalian DNA Polymerase δ−Proliferating Cell Nuclear Antigen−Template-Primer Complex: Molecular Characterization by Direct Binding†
Identifieur interne : 003E50 ( Main/Exploration ); précédent : 003E49; suivant : 003E51The Mammalian DNA Polymerase δ−Proliferating Cell Nuclear Antigen−Template-Primer Complex: Molecular Characterization by Direct Binding†
Auteurs : Maeve Mcconnell [États-Unis] ; Holly Miller [États-Unis] ; Dmitry Ju. Mozzherin [États-Unis] ; Aaron Quamina [États-Unis] ; Cheng-Keat Tan [États-Unis] ; Kathleen M. Downey [États-Unis] ; Paul A. Fisher [États-Unis]Source :
- Biochemistry [ 0006-2960 ] ; 1996.
Abstract
Three direct assays, polyacrylamide gel electrophoresis-band mobility shift, agarose gel electrophoresis-band mobility shift, and nitrocellulose filter binding, were established to study complexes formed among mammalian DNA polymerase δ (pol δ), proliferating cell nuclear antigen (PCNA), and synthetic oligonucleotide template-primers. In all contexts, complex formation requires simultaneous presence of pol δ, PCNA, and template-primer. Moreover, we showed in one such assay that the complex formed contains each molecular component. Nuclease protection experiments demonstrate that complex formation protects template from degradation by DNase I. The mass determined for the pol δ·PCNA· template-primer complex was about 267 kDa, consistent with the participation of one molecule of pol δ, two or three molecules of PCNA and one molecule of template-primer. PCNA alone behaved as a trimer (mass determined to be about 87 kDa). Complex could be manipulated enzymologically. Measurement of off rates demonstrates directly that PCNA stabilizes the pol δ·template-primer complex.
Url:
DOI: 10.1021/bi9530649
Affiliations:
Links toward previous steps (curation, corpus...)
- to stream Istex, to step Corpus: 000D74
- to stream Istex, to step Curation: 000D74
- to stream Istex, to step Checkpoint: 001586
- to stream Main, to step Merge: 003F05
- to stream Main, to step Curation: 003E50
Le document en format XML
<record><TEI wicri:istexFullTextTei="biblStruct"><teiHeader><fileDesc><titleStmt><title>The Mammalian DNA Polymerase δ−Proliferating Cell Nuclear Antigen−Template-Primer Complex: Molecular Characterization by Direct Binding†</title><author><name sortKey="Mcconnell, Maeve" sort="Mcconnell, Maeve" uniqKey="Mcconnell M" first="Maeve" last="Mcconnell">Maeve Mcconnell</name></author><author><name sortKey="Miller, Holly" sort="Miller, Holly" uniqKey="Miller H" first="Holly" last="Miller">Holly Miller</name></author><author><name sortKey="Mozzherin, Dmitry Ju" sort="Mozzherin, Dmitry Ju" uniqKey="Mozzherin D" first="Dmitry Ju." last="Mozzherin">Dmitry Ju. Mozzherin</name></author><author><name sortKey="Quamina, Aaron" sort="Quamina, Aaron" uniqKey="Quamina A" first="Aaron" last="Quamina">Aaron Quamina</name></author><author><name sortKey="Tan, Cheng Keat" sort="Tan, Cheng Keat" uniqKey="Tan C" first="Cheng-Keat" last="Tan">Cheng-Keat Tan</name></author><author><name sortKey="Downey, Kathleen M" sort="Downey, Kathleen M" uniqKey="Downey K" first="Kathleen M." last="Downey">Kathleen M. Downey</name></author><author><name sortKey="Fisher, Paul A" sort="Fisher, Paul A" uniqKey="Fisher P" first="Paul A." last="Fisher">Paul A. Fisher</name></author></titleStmt><publicationStmt><idno type="wicri:source">ISTEX</idno><idno type="RBID">ISTEX:1EF9F94BC65F34BED1E2365109FF1711414334DB</idno><date when="1996" year="1996">1996</date><idno type="doi">10.1021/bi9530649</idno><idno type="url">https://api.istex.fr/ark:/67375/TPS-S3LGXDJ3-R/fulltext.pdf</idno><idno type="wicri:Area/Istex/Corpus">000D74</idno><idno type="wicri:explorRef" wicri:stream="Istex" wicri:step="Corpus" wicri:corpus="ISTEX">000D74</idno><idno type="wicri:Area/Istex/Curation">000D74</idno><idno type="wicri:Area/Istex/Checkpoint">001586</idno><idno type="wicri:explorRef" wicri:stream="Istex" wicri:step="Checkpoint">001586</idno><idno type="wicri:doubleKey">0006-2960:1996:Mcconnell M:the:mammalian:dna</idno><idno type="wicri:Area/Main/Merge">003F05</idno><idno type="wicri:Area/Main/Curation">003E50</idno><idno type="wicri:Area/Main/Exploration">003E50</idno></publicationStmt><sourceDesc><biblStruct><analytic><title level="a" type="main">The Mammalian DNA Polymerase δ−Proliferating Cell Nuclear
Antigen−Template-Primer Complex: Molecular Characterization by Direct
Binding<ref type="bib" target="#bi9530649AF2"><hi rend="superscript">†</hi></ref></title><author><name sortKey="Mcconnell, Maeve" sort="Mcconnell, Maeve" uniqKey="Mcconnell M" first="Maeve" last="Mcconnell">Maeve Mcconnell</name><affiliation wicri:level="2"><country xml:lang="fr">États-Unis</country><placeName><region type="state">Floride</region></placeName><wicri:cityArea>Department of Pharmacological Sciences, University Medical Center, State University of New York at Stony Brook, StonyBrook, New York 11794-8651, and Departments of Medicine and Biochemistry, University of Miami School of Medicine, P.O.Box 016960, Miami</wicri:cityArea></affiliation><affiliation></affiliation></author><author><name sortKey="Miller, Holly" sort="Miller, Holly" uniqKey="Miller H" first="Holly" last="Miller">Holly Miller</name><affiliation wicri:level="2"><country xml:lang="fr">États-Unis</country><placeName><region type="state">Floride</region></placeName><wicri:cityArea>Department of Pharmacological Sciences, University Medical Center, State University of New York at Stony Brook, StonyBrook, New York 11794-8651, and Departments of Medicine and Biochemistry, University of Miami School of Medicine, P.O.Box 016960, Miami</wicri:cityArea></affiliation><affiliation></affiliation></author><author><name sortKey="Mozzherin, Dmitry Ju" sort="Mozzherin, Dmitry Ju" uniqKey="Mozzherin D" first="Dmitry Ju." last="Mozzherin">Dmitry Ju. Mozzherin</name><affiliation wicri:level="2"><country xml:lang="fr">États-Unis</country><placeName><region type="state">Floride</region></placeName><wicri:cityArea>Department of Pharmacological Sciences, University Medical Center, State University of New York at Stony Brook, StonyBrook, New York 11794-8651, and Departments of Medicine and Biochemistry, University of Miami School of Medicine, P.O.Box 016960, Miami</wicri:cityArea></affiliation><affiliation></affiliation></author><author><name sortKey="Quamina, Aaron" sort="Quamina, Aaron" uniqKey="Quamina A" first="Aaron" last="Quamina">Aaron Quamina</name><affiliation wicri:level="2"><country xml:lang="fr">États-Unis</country><placeName><region type="state">Floride</region></placeName><wicri:cityArea>Department of Pharmacological Sciences, University Medical Center, State University of New York at Stony Brook, StonyBrook, New York 11794-8651, and Departments of Medicine and Biochemistry, University of Miami School of Medicine, P.O.Box 016960, Miami</wicri:cityArea></affiliation><affiliation></affiliation></author><author><name sortKey="Tan, Cheng Keat" sort="Tan, Cheng Keat" uniqKey="Tan C" first="Cheng-Keat" last="Tan">Cheng-Keat Tan</name><affiliation wicri:level="2"><country xml:lang="fr">États-Unis</country><placeName><region type="state">Floride</region></placeName><wicri:cityArea>Department of Pharmacological Sciences, University Medical Center, State University of New York at Stony Brook, StonyBrook, New York 11794-8651, and Departments of Medicine and Biochemistry, University of Miami School of Medicine, P.O.Box 016960, Miami</wicri:cityArea></affiliation><affiliation></affiliation></author><author><name sortKey="Downey, Kathleen M" sort="Downey, Kathleen M" uniqKey="Downey K" first="Kathleen M." last="Downey">Kathleen M. Downey</name><affiliation wicri:level="2"><country xml:lang="fr">États-Unis</country><placeName><region type="state">Floride</region></placeName><wicri:cityArea>Department of Pharmacological Sciences, University Medical Center, State University of New York at Stony Brook, StonyBrook, New York 11794-8651, and Departments of Medicine and Biochemistry, University of Miami School of Medicine, P.O.Box 016960, Miami</wicri:cityArea></affiliation><affiliation></affiliation></author><author><name sortKey="Fisher, Paul A" sort="Fisher, Paul A" uniqKey="Fisher P" first="Paul A." last="Fisher">Paul A. Fisher</name><affiliation wicri:level="2"><country xml:lang="fr">États-Unis</country><placeName><region type="state">Floride</region></placeName><wicri:cityArea>Department of Pharmacological Sciences, University Medical Center, State University of New York at Stony Brook, StonyBrook, New York 11794-8651, and Departments of Medicine and Biochemistry, University of Miami School of Medicine, P.O.Box 016960, Miami</wicri:cityArea></affiliation><affiliation></affiliation><affiliation></affiliation></author></analytic><monogr></monogr><series><title level="j" type="main">Biochemistry</title><title level="j" type="abbrev">Biochemistry</title><idno type="ISSN">0006-2960</idno><idno type="eISSN">1520-4995</idno><imprint><publisher>American Chemical Society</publisher><date type="e-published">1996</date><date type="published">1996</date><biblScope unit="vol">35</biblScope><biblScope unit="issue">25</biblScope><biblScope unit="page" from="8268">8268</biblScope><biblScope unit="page" to="8274">8274</biblScope></imprint><idno type="ISSN">0006-2960</idno></series></biblStruct></sourceDesc><seriesStmt><idno type="ISSN">0006-2960</idno></seriesStmt></fileDesc><profileDesc><textClass></textClass></profileDesc></teiHeader><front><div type="abstract">Three direct assays, polyacrylamide gel electrophoresis-band mobility shift, agarose gel electrophoresis-band mobility shift, and nitrocellulose filter binding, were established to study complexes formed among mammalian DNA polymerase δ (pol δ), proliferating cell nuclear antigen (PCNA), and synthetic oligonucleotide template-primers. In all contexts, complex formation requires simultaneous presence of pol δ, PCNA, and template-primer. Moreover, we showed in one such assay that the complex formed contains each molecular component. Nuclease protection experiments demonstrate that complex formation protects template from degradation by DNase I. The mass determined for the pol δ·PCNA· template-primer complex was about 267 kDa, consistent with the participation of one molecule of pol δ, two or three molecules of PCNA and one molecule of template-primer. PCNA alone behaved as a trimer (mass determined to be about 87 kDa). Complex could be manipulated enzymologically. Measurement of off rates demonstrates directly that PCNA stabilizes the pol δ·template-primer complex.</div></front></TEI><affiliations><list><country><li>États-Unis</li></country><region><li>Floride</li></region></list><tree><country name="États-Unis"><region name="Floride"><name sortKey="Mcconnell, Maeve" sort="Mcconnell, Maeve" uniqKey="Mcconnell M" first="Maeve" last="Mcconnell">Maeve Mcconnell</name></region><name sortKey="Downey, Kathleen M" sort="Downey, Kathleen M" uniqKey="Downey K" first="Kathleen M." last="Downey">Kathleen M. Downey</name><name sortKey="Fisher, Paul A" sort="Fisher, Paul A" uniqKey="Fisher P" first="Paul A." last="Fisher">Paul A. Fisher</name><name sortKey="Miller, Holly" sort="Miller, Holly" uniqKey="Miller H" first="Holly" last="Miller">Holly Miller</name><name sortKey="Mozzherin, Dmitry Ju" sort="Mozzherin, Dmitry Ju" uniqKey="Mozzherin D" first="Dmitry Ju." last="Mozzherin">Dmitry Ju. Mozzherin</name><name sortKey="Quamina, Aaron" sort="Quamina, Aaron" uniqKey="Quamina A" first="Aaron" last="Quamina">Aaron Quamina</name><name sortKey="Tan, Cheng Keat" sort="Tan, Cheng Keat" uniqKey="Tan C" first="Cheng-Keat" last="Tan">Cheng-Keat Tan</name></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
EXPLOR_STEP=$WICRI_ROOT/Sante/explor/MersV1/Data/Main/Exploration
HfdSelect -h $EXPLOR_STEP/biblio.hfd -nk 003E50 | SxmlIndent | more
Ou
HfdSelect -h $EXPLOR_AREA/Data/Main/Exploration/biblio.hfd -nk 003E50 | SxmlIndent | more
Pour mettre un lien sur cette page dans le réseau Wicri
{{Explor lien
|wiki= Sante
|area= MersV1
|flux= Main
|étape= Exploration
|type= RBID
|clé= ISTEX:1EF9F94BC65F34BED1E2365109FF1711414334DB
|texte= The Mammalian DNA Polymerase δ−Proliferating Cell Nuclear Antigen−Template-Primer Complex: Molecular Characterization by Direct Binding†
}}
| This area was generated with Dilib version V0.6.33. |  |