Stoichiometric complex formation by proliferating cell nuclear antigen (PCNA) and its interacting protein: Purification and crystallization of the DNA polymerase and PCNA monomer mutant complex from Pyrococcus furiosus

Hirokazu Nishida, Shigeki Matsumiya, Daisuke Tsuchiya, Yoshizumi Ishino, Kosuke Morikawa

Research output: Contribution to journalArticlepeer-review

3 Citations (Scopus)

Abstract

Replicative DNA polymerase interacts with processivity factors, the β-subunit of DNA polymerase III or proliferating cell nuclear antigen (PCNA), in order to function with a long template DNA. The archaeal replicative DNA polymerase from Pyrococcus furiosus interacts with PCNA via its PCNA-interacting protein (PIP) motif at the C-terminus. The PCNA homotrimeric ring contains one PIP interacting site on each monomer and since the ring can accommodate up to three molecules simultaneously, formation of a stable stoichiometric complex of PCNA with its interacting protein has been difficult to control in vitro. A stable complex of the DNA polymerase with PCNA, using a PCNA monomer mutant, has been purified and crystallized. The best ordered crystal diffracted to 3.0 Å resolution using synchrotron radiation. The crystals belong to space group P21212, with unit-cell parameters a = 225.3, b = 123.3, c = 91.3 Å.

Original languageEnglish
Pages (from-to)253-256
Number of pages4
JournalActa Crystallographica Section F: Structural Biology and Crystallization Communications
Volume62
Issue number3
DOIs
Publication statusPublished - 2006

All Science Journal Classification (ASJC) codes

  • Biophysics
  • Structural Biology
  • Biochemistry
  • Genetics
  • Condensed Matter Physics

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