A single amino acid substitution in the DNA-binding domain of Aeropyrum pernix DNA ligase impairs its interaction with proliferating cell nuclear antigen

Shinichi Kiyonari, Toru Kamigochi, Yoshizumi Ishino

Research output: Contribution to journalArticle

8 Citations (Scopus)

Abstract

Proliferating cell nuclear antigen (PCNA) is known as a DNA sliding clamp that acts as a platform for the assembly of enzymes involved in DNA replication and repair. Previously, it was reported that a crenarchaeal PCNA formed a heterotrimeric structure, and that each PCNA subunit has distinct binding specificity to PCNA-binding proteins. Here we describe the PCNA-binding properties of a DNA ligase from the hyperthermophilic crenarchaeon Aeropyrum pernix K1. Based on our findings on the Pyrococcus furiosus DNA ligase-PCNA interaction, we predicted that the aromatic residue, Phe132, in the DNA-binding domain of A. pernix DNA ligase (ApeLig) would play a critical role in binding to A. pernix PCNA (ApePCNA). Surface plasmon resonance analyses revealed that the ApeLig F132A mutant does not interact with an immobilized subunit of ApePCNA. Furthermore, we could not detect any stimulation of the ligation activity of the ApeLig F132A protein by ApePCNA in vitro. These results indicated that the phenylalanine, which is located in our predicted PCNA-binding region in ApeLig, has a critical role for the physical and functional interaction with ApePCNA.

Original languageEnglish
Pages (from-to)675-684
Number of pages10
JournalExtremophiles
Volume11
Issue number5
DOIs
Publication statusPublished - Sep 1 2007

Fingerprint

Aeropyrum
DNA Ligases
Proliferating Cell Nuclear Antigen
Amino Acid Substitution
DNA
Pyrococcus furiosus
Surface Plasmon Resonance
Staphylococcal Protein A
Phenylalanine
DNA Replication
DNA Repair

All Science Journal Classification (ASJC) codes

  • Microbiology
  • Molecular Medicine

Cite this

A single amino acid substitution in the DNA-binding domain of Aeropyrum pernix DNA ligase impairs its interaction with proliferating cell nuclear antigen. / Kiyonari, Shinichi; Kamigochi, Toru; Ishino, Yoshizumi.

In: Extremophiles, Vol. 11, No. 5, 01.09.2007, p. 675-684.

Research output: Contribution to journalArticle

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