Crystal structure of a biologically functional form of PriB from Escherichia coli reveals a potential single-stranded DNA-binding site

Seijiro Shioi, Toyoyuki Ose, Katsumi Maenaka, Mitsunori Shiroishi, Yoshito Abe, Daisuke Kohda, Tsutomu Katayama, Tadashi Ueda

Research output: Contribution to journalArticle

24 Citations (Scopus)

Abstract

PriB is not only an essential protein necessary for the replication restart on the collapsed and disintegrated replication fork, but also an important protein for assembling of primosome onto ΦX174 genomic DNA during replication initiation. Here we report a 2.0-Å-resolution X-ray structure of a biologically functional form of PriB from Escherichia coli. The crystal structure revealed that despite a low level of primary sequence identity, the PriB monomer, as well as the dimeric form, are structurally identical to the N-terminal DNA-binding domain of the single-stranded DNA-binding protein (SSB) from Escherichia coli, which possesses an oligonucleotides-binding-fold. The oligonucleotide-PriB complex model based on the oligonucleotides-SSB complex structure suggested that PriB had a DNA-binding pocket conserved in SSB from Escherichia coli and might bind to single-stranded DNA in the manner of SSB. Furthermore, surface plasmon resonance analysis and fluorescence measurements demonstrated that PriB binds single-stranded DNA with high affinity, by involving tryptophan residue. The significance of these results with respect to the functional role of PriB in the assembly of primosome is discussed.

Original languageEnglish
Pages (from-to)766-776
Number of pages11
JournalBiochemical and Biophysical Research Communications
Volume326
Issue number4
DOIs
Publication statusPublished - Jan 28 2005

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Single-Stranded DNA
Oligonucleotides
Escherichia coli
Crystal structure
Binding Sites
DNA
Surface Plasmon Resonance
DNA-Binding Proteins
Surface plasmon resonance
DNA Replication
Tryptophan
Proteins
Monomers
Fluorescence
X-Rays
X rays

All Science Journal Classification (ASJC) codes

  • Biophysics
  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Crystal structure of a biologically functional form of PriB from Escherichia coli reveals a potential single-stranded DNA-binding site. / Shioi, Seijiro; Ose, Toyoyuki; Maenaka, Katsumi; Shiroishi, Mitsunori; Abe, Yoshito; Kohda, Daisuke; Katayama, Tsutomu; Ueda, Tadashi.

In: Biochemical and Biophysical Research Communications, Vol. 326, No. 4, 28.01.2005, p. 766-776.

Research output: Contribution to journalArticle

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AU - Abe, Yoshito

AU - Kohda, Daisuke

AU - Katayama, Tsutomu

AU - Ueda, Tadashi

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