Basic and aromatic residues in the C-terminal domain of PriC are involved in ssDNA and SSB binding

Takahiko Aramaki, Yoshito Abe, Kaori Furutani, Tsutomu Katayama, Tadashi Ueda

Research output: Contribution to journalArticle

7 Citations (Scopus)

Abstract

In bacterial organisms, the oriC-independent primosome plays an essential role in replication restart after dissociation of the replication DNA-protein complex following DNA damage. PriC is a key protein component in the oriC-independent replication restart primosome. Our previous study suggested that PriC was divided into an N-terminal domain and a C-terminal domain, with the latter domain being the major contributor to single-stranded DNA (ssDNA) binding capacity. In this study, we prepared several PriC mutants in which basic and aromatic amino acid residues were mutated to alanine. Five of these residues, Arg107, Lys111, Phe118, Arg121 and Lys165 in the C-terminal domain, were shown to be involved in ssDNA binding. Moreover, we evaluated the binding of the PriC mutants to the ssDNA-binding protein (SSB) complex. Five residues, Phe118, Arg121, Arg129, Tyr152 and Arg155 in the C-terminal domain of PriC, were shown to be involved in SSB binding in the presence of ssDNA. On the basis of these results, we propose a structural model of the C-terminal domain of PriC and discuss how the interactions of PriC with SSB and ssDNA may contribute to the regulation of PriC-dependent replication restart.

Original languageEnglish
Pages (from-to)529-537
Number of pages9
JournalJournal of Biochemistry
Volume157
Issue number6
DOIs
Publication statusPublished - Jan 1 2015

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Single-Stranded DNA
DNA-Binding Proteins
Protein Binding
Basic Amino Acids
Aromatic Amino Acids
Structural Models
DNA
DNA Replication
Alanine
DNA Damage
Carrier Proteins
Proteins

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Molecular Biology

Cite this

Basic and aromatic residues in the C-terminal domain of PriC are involved in ssDNA and SSB binding. / Aramaki, Takahiko; Abe, Yoshito; Furutani, Kaori; Katayama, Tsutomu; Ueda, Tadashi.

In: Journal of Biochemistry, Vol. 157, No. 6, 01.01.2015, p. 529-537.

Research output: Contribution to journalArticle

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AU - Abe, Yoshito

AU - Furutani, Kaori

AU - Katayama, Tsutomu

AU - Ueda, Tadashi

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