Intermolecular ion pairs maintain the toroidal structure of Pyrococcus furiosus PCNA

Shigeki Matsumiya, Sonoko Ishino, Yoshizumi Ishino, Kosuke Morikawa

Research output: Contribution to journalArticle

30 Citations (Scopus)

Abstract

Two mutant proliferating cell nuclear antigens from the hyperthermophilic archaeon Pyrococcus furiosus, PfuPCNA(D143A) and PfuPCNA(D143A/D147A), were prepared by site-specific mutagenesis. The results from gel filtration showed that mutations at D143 and D147 drastically affect the stability of the trimeric structure of PfuPCNA. The PfuPCNA(D143A) still retained the activity to stimulate the DNA polymerase reaction, but PfuPCNA(D143A/D147A) lost the activity. Crystal structures of the mutant PfuPCNAs were determined. Although the wild-type PCNA forms a toroidal trimer with intermolecular hydrogen bonds between the N- and C-terminal domains, the mutant PfuPCNAs exist as V-shaped dimers through intermolecular hydrogen bonds between the two C-terminal domains in the crystal. Because the mutated residues are involved in the intermolecular ion pairs through their side chains in the wild-type PfuPCNA, these ion pairs seem to play a key role in maintaining the toroidal structure of the PfuPCNA trimer. The comparison of the crystal structures revealed intriguing conformational flexibility of each domain in the PfuPCNA subunit. This structural versatility of PCNA may be involved in the mechanisms for ring opening and closing.

Original languageEnglish
Pages (from-to)823-831
Number of pages9
JournalProtein Science
Volume12
Issue number4
DOIs
Publication statusPublished - Apr 1 2003

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Pyrococcus furiosus
Proliferating Cell Nuclear Antigen
Ions
Hydrogen
Hydrogen bonds
Crystal structure
Mutagenesis
Archaea
DNA-Directed DNA Polymerase
Site-Directed Mutagenesis
Dimers
Gel Chromatography
Gels
Mutation
Crystals

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Molecular Biology

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Intermolecular ion pairs maintain the toroidal structure of Pyrococcus furiosus PCNA. / Matsumiya, Shigeki; Ishino, Sonoko; Ishino, Yoshizumi; Morikawa, Kosuke.

In: Protein Science, Vol. 12, No. 4, 01.04.2003, p. 823-831.

Research output: Contribution to journalArticle

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