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

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72 Citations (Scopus)

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.

Original languageEnglish
Pages (from-to)40362-40367
Number of pages6
JournalJournal of Biological Chemistry
Volume277
Issue number43
DOIs
Publication statusPublished - Oct 25 2002
Externally publishedYes

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All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Molecular Biology
  • Cell Biology

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