DNA polymerase lambda directly binds to proliferating cell nuclear antigen through its confined C-terminal region.
Identifieur interne : 000349 ( Ncbi/Checkpoint ); précédent : 000348; suivant : 000350DNA polymerase lambda directly binds to proliferating cell nuclear antigen through its confined C-terminal region.
Auteurs : Noriko Shimazaki [Japon] ; Takaya Yazaki ; Takashi Kubota ; Asami Sato ; Ayako Nakamura ; Shunsuke Kurei ; Shingo Toji ; Katsuyuki Tamai ; Osamu KoiwaiSource :
- Genes to cells : devoted to molecular & cellular mechanisms [ 1356-9597 ] ; 2005.
Descripteurs français
- KwdFr :
- Antigène nucléaire de prolifération cellulaire (génétique), Antigène nucléaire de prolifération cellulaire (métabolisme), DNA nucleotidylexotransferase (métabolisme), DNA polymerase beta (génétique), DNA polymerase beta (métabolisme), Humains, Immunoprécipitation, Liaison aux protéines, Microscopie de fluorescence, Mutagenèse dirigée, Mutation, Noyau de la cellule (métabolisme), Protéines à fluorescence verte (métabolisme), Réplication de l'ADN, Sites de fixation, Structure tertiaire des protéines, Techniques in vitro.
- MESH :
- génétique : Antigène nucléaire de prolifération cellulaire, DNA polymerase beta.
- métabolisme : Antigène nucléaire de prolifération cellulaire, DNA nucleotidylexotransferase, DNA polymerase beta, Noyau de la cellule, Protéines à fluorescence verte.
- Humains, Immunoprécipitation, Liaison aux protéines, Microscopie de fluorescence, Mutagenèse dirigée, Mutation, Réplication de l'ADN, Sites de fixation, Structure tertiaire des protéines, Techniques in vitro.
English descriptors
- KwdEn :
- Binding Sites, Cell Nucleus (metabolism), DNA Nucleotidylexotransferase (metabolism), DNA Polymerase beta (genetics), DNA Polymerase beta (metabolism), DNA Replication, Green Fluorescent Proteins (metabolism), Humans, Immunoprecipitation, In Vitro Techniques, Microscopy, Fluorescence, Mutagenesis, Site-Directed, Mutation, Proliferating Cell Nuclear Antigen (genetics), Proliferating Cell Nuclear Antigen (metabolism), Protein Binding, Protein Structure, Tertiary.
- MESH :
- chemical , genetics : DNA Polymerase beta, Proliferating Cell Nuclear Antigen.
- chemical , metabolism : DNA Nucleotidylexotransferase, DNA Polymerase beta, Green Fluorescent Proteins, Proliferating Cell Nuclear Antigen.
- metabolism : Cell Nucleus.
- Binding Sites, DNA Replication, Humans, Immunoprecipitation, In Vitro Techniques, Microscopy, Fluorescence, Mutagenesis, Site-Directed, Mutation, Protein Binding, Protein Structure, Tertiary.
Abstract
DNA polymerase lambda (Pol lambda) was recently identified as a new member of the family X of DNA polymerases. Here, we show that Pol lambda directly binds to proliferating cell nuclear antigen (PCNA), an auxiliary protein for DNA replication and repair enzymes, both in vitro and in vivo. A pull-down assay using deletion mutants of Pol lambda showed that the confined C-terminal region of Pol lambda directly binds to PCNA. Furthermore, a synthetic peptide of 20-mers derived from the C-terminal region of Pol lambda competes with full-length Pol lambda for binding to PCNA. The residues between amino acids 518 and 537 of Pol lambda are required for binding to PCNA, and are different from the consensus PCNA interacting motif (PIM). Pol lambda associates with PCNA in vivo by immunoprecipitation analysis and EGFP-tagged Pol lambda co-localizes with PCNA as spots within a nucleus using fluorescent microscopy. Through direct binding, PCNA suppressed the distributive nucleotidyltransferase activity of Pol lambda. Pol micro, which also belongs to the family X of DNA polymerases, binds to PCNA by a pivotal amino acid residue.
DOI: 10.1111/j.1365-2443.2005.00868.x
PubMed: 15966901
Affiliations:
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pubmed:15966901Le document en format XML
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<author><name sortKey="Shimazaki, Noriko" sort="Shimazaki, Noriko" uniqKey="Shimazaki N" first="Noriko" last="Shimazaki">Noriko Shimazaki</name>
<affiliation wicri:level="1"><nlm:affiliation>Department of Applied Biological Science, Faculty of Science and Technology, Tokyo University of Science, Noda, Chiba 278-8510, Japan. snoriko@rs.noda.tus.ac.jp</nlm:affiliation>
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<profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Binding Sites</term>
<term>Cell Nucleus (metabolism)</term>
<term>DNA Nucleotidylexotransferase (metabolism)</term>
<term>DNA Polymerase beta (genetics)</term>
<term>DNA Polymerase beta (metabolism)</term>
<term>DNA Replication</term>
<term>Green Fluorescent Proteins (metabolism)</term>
<term>Humans</term>
<term>Immunoprecipitation</term>
<term>In Vitro Techniques</term>
<term>Microscopy, Fluorescence</term>
<term>Mutagenesis, Site-Directed</term>
<term>Mutation</term>
<term>Proliferating Cell Nuclear Antigen (genetics)</term>
<term>Proliferating Cell Nuclear Antigen (metabolism)</term>
<term>Protein Binding</term>
<term>Protein Structure, Tertiary</term>
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<keywords scheme="KwdFr" xml:lang="fr"><term>Antigène nucléaire de prolifération cellulaire (génétique)</term>
<term>Antigène nucléaire de prolifération cellulaire (métabolisme)</term>
<term>DNA nucleotidylexotransferase (métabolisme)</term>
<term>DNA polymerase beta (génétique)</term>
<term>DNA polymerase beta (métabolisme)</term>
<term>Humains</term>
<term>Immunoprécipitation</term>
<term>Liaison aux protéines</term>
<term>Microscopie de fluorescence</term>
<term>Mutagenèse dirigée</term>
<term>Mutation</term>
<term>Noyau de la cellule (métabolisme)</term>
<term>Protéines à fluorescence verte (métabolisme)</term>
<term>Réplication de l'ADN</term>
<term>Sites de fixation</term>
<term>Structure tertiaire des protéines</term>
<term>Techniques in vitro</term>
</keywords>
<keywords scheme="MESH" type="chemical" qualifier="genetics" xml:lang="en"><term>DNA Polymerase beta</term>
<term>Proliferating Cell Nuclear Antigen</term>
</keywords>
<keywords scheme="MESH" type="chemical" qualifier="metabolism" xml:lang="en"><term>DNA Nucleotidylexotransferase</term>
<term>DNA Polymerase beta</term>
<term>Green Fluorescent Proteins</term>
<term>Proliferating Cell Nuclear Antigen</term>
</keywords>
<keywords scheme="MESH" qualifier="génétique" xml:lang="fr"><term>Antigène nucléaire de prolifération cellulaire</term>
<term>DNA polymerase beta</term>
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<term>DNA nucleotidylexotransferase</term>
<term>DNA polymerase beta</term>
<term>Noyau de la cellule</term>
<term>Protéines à fluorescence verte</term>
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<keywords scheme="MESH" xml:lang="en"><term>Binding Sites</term>
<term>DNA Replication</term>
<term>Humans</term>
<term>Immunoprecipitation</term>
<term>In Vitro Techniques</term>
<term>Microscopy, Fluorescence</term>
<term>Mutagenesis, Site-Directed</term>
<term>Mutation</term>
<term>Protein Binding</term>
<term>Protein Structure, Tertiary</term>
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<keywords scheme="MESH" xml:lang="fr"><term>Humains</term>
<term>Immunoprécipitation</term>
<term>Liaison aux protéines</term>
<term>Microscopie de fluorescence</term>
<term>Mutagenèse dirigée</term>
<term>Mutation</term>
<term>Réplication de l'ADN</term>
<term>Sites de fixation</term>
<term>Structure tertiaire des protéines</term>
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<front><div type="abstract" xml:lang="en">DNA polymerase lambda (Pol lambda) was recently identified as a new member of the family X of DNA polymerases. Here, we show that Pol lambda directly binds to proliferating cell nuclear antigen (PCNA), an auxiliary protein for DNA replication and repair enzymes, both in vitro and in vivo. A pull-down assay using deletion mutants of Pol lambda showed that the confined C-terminal region of Pol lambda directly binds to PCNA. Furthermore, a synthetic peptide of 20-mers derived from the C-terminal region of Pol lambda competes with full-length Pol lambda for binding to PCNA. The residues between amino acids 518 and 537 of Pol lambda are required for binding to PCNA, and are different from the consensus PCNA interacting motif (PIM). Pol lambda associates with PCNA in vivo by immunoprecipitation analysis and EGFP-tagged Pol lambda co-localizes with PCNA as spots within a nucleus using fluorescent microscopy. Through direct binding, PCNA suppressed the distributive nucleotidyltransferase activity of Pol lambda. Pol micro, which also belongs to the family X of DNA polymerases, binds to PCNA by a pivotal amino acid residue.</div>
</front>
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<tree><noCountry><name sortKey="Koiwai, Osamu" sort="Koiwai, Osamu" uniqKey="Koiwai O" first="Osamu" last="Koiwai">Osamu Koiwai</name>
<name sortKey="Kubota, Takashi" sort="Kubota, Takashi" uniqKey="Kubota T" first="Takashi" last="Kubota">Takashi Kubota</name>
<name sortKey="Kurei, Shunsuke" sort="Kurei, Shunsuke" uniqKey="Kurei S" first="Shunsuke" last="Kurei">Shunsuke Kurei</name>
<name sortKey="Nakamura, Ayako" sort="Nakamura, Ayako" uniqKey="Nakamura A" first="Ayako" last="Nakamura">Ayako Nakamura</name>
<name sortKey="Sato, Asami" sort="Sato, Asami" uniqKey="Sato A" first="Asami" last="Sato">Asami Sato</name>
<name sortKey="Tamai, Katsuyuki" sort="Tamai, Katsuyuki" uniqKey="Tamai K" first="Katsuyuki" last="Tamai">Katsuyuki Tamai</name>
<name sortKey="Toji, Shingo" sort="Toji, Shingo" uniqKey="Toji S" first="Shingo" last="Toji">Shingo Toji</name>
<name sortKey="Yazaki, Takaya" sort="Yazaki, Takaya" uniqKey="Yazaki T" first="Takaya" last="Yazaki">Takaya Yazaki</name>
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<country name="Japon"><noRegion><name sortKey="Shimazaki, Noriko" sort="Shimazaki, Noriko" uniqKey="Shimazaki N" first="Noriko" last="Shimazaki">Noriko Shimazaki</name>
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