PCNA clamp facilitates action of DNA cytosine methyltransferase 1 on hemimethylated DNA

Tetsuo Iida, Isao Suetake, Shoji Tajima, Hiroshi Morioka, Satoshi Ohta, Chikashi Obuse, Toshiki Tsurimoto

研究成果: ジャーナルへの寄稿記事

86 引用 (Scopus)

抄録

Background: Proliferating cell nuclear antigen (PCNA) is a ring-shaped protein known as a processivity factor of DNA polymerase δ. In addition to this role, PCNA interacts with a number of other proteins to increase their local concentration at replicated DNA sites. DNA cytosine methyltransferase 1 (Dnmt1), which preserves epigenetic signals by completing the methylation of hemimethylated DNA after DNA replication, has been indicated as one of these PCNA binding proteins by a previous work. However, the molecular mechanisms and functional significance of their association have not yet been studied. Results: Dnmt1 can be readily isolated from nuclear extracts by PCNA affinity chromatography. Studies of the interactions between the two proteins demonstrate that the N-terminal region of Dnmt1, which contains a typical PCNA binding motif, has core PCNA binding activity, and that the remaining portion of the protein exerts a negative influence on the interaction of Dnmt1 with PCNA.The affinity of Dnmt1 for DNA is much higher for DNA bound by PCNA than for free DNA. Furthermore, DNA methylation assays with hemimethylated DNA as a substrate revealed that PCNA clamp-bound DNA is methylated more efficiently by Dnmt1 than is free DNA. Conclusion: These results provide the first biochemical evidence that physical interactions between PCNA and Dnmt1 facilitate the methylation of newly neplicated DNA, on which PCNA remains associated as a functional clamp.

元の言語英語
ページ(範囲)997-1007
ページ数11
ジャーナルGenes to Cells
7
発行部数10
DOI
出版物ステータス出版済み - 10 1 2002
外部発表Yes

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Cytosine
Proliferating Cell Nuclear Antigen
Methyltransferases
DNA
DNA Methylation
Proteins
DNA-Directed DNA Polymerase
DNA Replication
Affinity Chromatography
Epigenomics
Methylation

All Science Journal Classification (ASJC) codes

  • Genetics
  • Cell Biology

これを引用

PCNA clamp facilitates action of DNA cytosine methyltransferase 1 on hemimethylated DNA. / Iida, Tetsuo; Suetake, Isao; Tajima, Shoji; Morioka, Hiroshi; Ohta, Satoshi; Obuse, Chikashi; Tsurimoto, Toshiki.

:: Genes to Cells, 巻 7, 番号 10, 01.10.2002, p. 997-1007.

研究成果: ジャーナルへの寄稿記事

Iida, Tetsuo ; Suetake, Isao ; Tajima, Shoji ; Morioka, Hiroshi ; Ohta, Satoshi ; Obuse, Chikashi ; Tsurimoto, Toshiki. / PCNA clamp facilitates action of DNA cytosine methyltransferase 1 on hemimethylated DNA. :: Genes to Cells. 2002 ; 巻 7, 番号 10. pp. 997-1007.
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abstract = "Background: Proliferating cell nuclear antigen (PCNA) is a ring-shaped protein known as a processivity factor of DNA polymerase δ. In addition to this role, PCNA interacts with a number of other proteins to increase their local concentration at replicated DNA sites. DNA cytosine methyltransferase 1 (Dnmt1), which preserves epigenetic signals by completing the methylation of hemimethylated DNA after DNA replication, has been indicated as one of these PCNA binding proteins by a previous work. However, the molecular mechanisms and functional significance of their association have not yet been studied. Results: Dnmt1 can be readily isolated from nuclear extracts by PCNA affinity chromatography. Studies of the interactions between the two proteins demonstrate that the N-terminal region of Dnmt1, which contains a typical PCNA binding motif, has core PCNA binding activity, and that the remaining portion of the protein exerts a negative influence on the interaction of Dnmt1 with PCNA.The affinity of Dnmt1 for DNA is much higher for DNA bound by PCNA than for free DNA. Furthermore, DNA methylation assays with hemimethylated DNA as a substrate revealed that PCNA clamp-bound DNA is methylated more efficiently by Dnmt1 than is free DNA. Conclusion: These results provide the first biochemical evidence that physical interactions between PCNA and Dnmt1 facilitate the methylation of newly neplicated DNA, on which PCNA remains associated as a functional clamp.",
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