A proteomics approach to identify proliferating cell nuclear antigen (PCNA)-binding proteins in human cell lysates: Identification of the human CHL12/RFCs2-5 complex as a novel PCNA-binding protein

Satoshi Ohta, Yasushi Shiomi, Katsunori Sugimoto, Chikashi Obuse, Toshiki Tsurimoto

研究成果: ジャーナルへの寄稿記事

69 引用 (Scopus)

抄録

Proliferating cell nuclear antigen (PCNA), a eukaryotic DNA replication factor, functions not only as a processivity factor for DNA polymerase 6 but also as a binding partner for multiple other factors. To understand its broad significance, we have carried out systematic studies of PCNA-binding proteins by a combination of affinity chromatography and mass spectrometric analyses. We detected more than 20 specific protein bands of various intensities in fractions bound to PCNA-fixed resin from human cell lysates and determined their peptide sequences by liquid chromatography and tandem mass spectrometry. A search with human protein data bases identified 12 reported PCNA-binding proteins from both cytoplasmic (S100 lysate) and nuclear extracts with substantial significance and four more solely from the nuclear preparation. CHL12, a factor involved in check-point response and chromosome cohesion, was a novel example found in both lysates. Further studies with recombinant proteins demonstrated that CHL12 and small subunits of replication factor C form a complex that interacts with PCNA.

元の言語英語
ページ(範囲)40362-40367
ページ数6
ジャーナルJournal of Biological Chemistry
277
発行部数43
DOI
出版物ステータス出版済み - 10 25 2002

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Forensic Anthropology
Proliferating Cell Nuclear Antigen
Proteomics
Carrier Proteins
Cells
Replication Protein C
Affinity chromatography
Sexual Partners
Liquid chromatography
DNA-Directed DNA Polymerase
Chromosomes
Tandem Mass Spectrometry
DNA Replication
Affinity Chromatography
Recombinant Proteins
Liquid Chromatography
Mass spectrometry
Proteins
Resins
Databases

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Molecular Biology
  • Cell Biology

これを引用

A proteomics approach to identify proliferating cell nuclear antigen (PCNA)-binding proteins in human cell lysates : Identification of the human CHL12/RFCs2-5 complex as a novel PCNA-binding protein. / Ohta, Satoshi; Shiomi, Yasushi; Sugimoto, Katsunori; Obuse, Chikashi; Tsurimoto, Toshiki.

:: Journal of Biological Chemistry, 巻 277, 番号 43, 25.10.2002, p. 40362-40367.

研究成果: ジャーナルへの寄稿記事

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abstract = "Proliferating cell nuclear antigen (PCNA), a eukaryotic DNA replication factor, functions not only as a processivity factor for DNA polymerase 6 but also as a binding partner for multiple other factors. To understand its broad significance, we have carried out systematic studies of PCNA-binding proteins by a combination of affinity chromatography and mass spectrometric analyses. We detected more than 20 specific protein bands of various intensities in fractions bound to PCNA-fixed resin from human cell lysates and determined their peptide sequences by liquid chromatography and tandem mass spectrometry. A search with human protein data bases identified 12 reported PCNA-binding proteins from both cytoplasmic (S100 lysate) and nuclear extracts with substantial significance and four more solely from the nuclear preparation. CHL12, a factor involved in check-point response and chromosome cohesion, was a novel example found in both lysates. Further studies with recombinant proteins demonstrated that CHL12 and small subunits of replication factor C form a complex that interacts with PCNA.",
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