Crystallization of proliferating cell nuclear antigen (PCNA) from saccharomyces cerevisiae

Talluru S.R. Krishna, David Fenyö, Xiang Peng Kong, Sonja Gary, Brian T. Chait, Peter Burgers, John Kuriyan

Research output: Contribution to journalArticle

35 Scopus citations

Abstract

Proliferating cell nuclear antigen (PCNA) is the component of the chromosomal DNA replication machinery in eukaryotic cells that confers high processivity upon DNA polymerase γ and ϵ. It has been proposed that PCNA functions by forming a trimeric complex with a ring-like structure through which DNA is threaded. PCNA from the yeast Saccharomyces cerevisiae has been crystallized in a cubic space group (P213, a=121.1 Å). Unexpectedly, a mercury derivative of PCNA yields crystals that diffract significantly better than crystals of the unmodified protein (2.4 Å and 3.0 Å resolution, respectively). Mass spectrometry reveals that the derivative results from the addition of two mercury atoms to the protein. Although crystals of the mercurated protein show evidence of nonisomorphism, the anomalous diffraction signal is strong and phases may be determined by multi-wavelength anomalous diffraction (MAD phasing).

Original languageEnglish
Pages (from-to)265-268
Number of pages4
JournalJournal of Molecular Biology
Volume241
Issue number2
DOIs
StatePublished - Aug 11 1994

Keywords

  • Crystallography
  • DNA polymerase
  • DNA replication
  • Processivity

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