Conserved hydrophobic amino acid residues in the N-terminal region of DnaA protein are involved in DnaA-DnaA interaction

Shinji Mima, Masaki Makise, Motohiro Koterasawa, Tomofusa Tsuchiya, Tohru Mizushima

Research output: Contribution to journalArticle

3 Citations (Scopus)

Abstract

We previously reported that a leucine-zipper-like structure (I26, L33 and L40) located in the N-terminal region of DnaA is essential for the duplex opening at oriC by DnaA. In this study, we focused on three other conserved hydrophobic amino acid residues, L3, L10 and L17, and examined the function of DnaA proteins mutated in these amino acid residues. DnaA427 (L17S) and DnaA413 (L3S, L10S and L17S) were inactive for oriC DNA replication both in vitro and in vivo. Although these mutant DnaA proteins maintained their binding activities for both ATP and oriC, they were unable to induce the opening of duplex DNA at oriC. Glutathione-S-transferase (GST)-fused wild-type DnaA interacted with wild-type DnaA but not with DnaA427 and DnaA413. Based on these results, we propose that conserved hydrophobic amino acid residues in the N-terminal region of DnaA are involved in DnaA oligomerization, in which DnaA-DnaA interaction is required.

Original languageEnglish
Pages (from-to)881-887
Number of pages7
JournalBiochemical Journal
Volume365
Issue number3
DOIs
Publication statusPublished - 2002 Aug 1
Externally publishedYes

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Amino Acids
dextran - saline drug combination
Leucine Zippers
Oligomerization
Proteins
DNA
Mutant Proteins
Glutathione Transferase
DNA Replication
Adenosine Triphosphate
In Vitro Techniques

Keywords

  • DNA replication
  • DnaA oligomerization
  • Hydrophobic amino acid residues
  • N-terminal

ASJC Scopus subject areas

  • Biochemistry

Cite this

Conserved hydrophobic amino acid residues in the N-terminal region of DnaA protein are involved in DnaA-DnaA interaction. / Mima, Shinji; Makise, Masaki; Koterasawa, Motohiro; Tsuchiya, Tomofusa; Mizushima, Tohru.

In: Biochemical Journal, Vol. 365, No. 3, 01.08.2002, p. 881-887.

Research output: Contribution to journalArticle

Mima, Shinji ; Makise, Masaki ; Koterasawa, Motohiro ; Tsuchiya, Tomofusa ; Mizushima, Tohru. / Conserved hydrophobic amino acid residues in the N-terminal region of DnaA protein are involved in DnaA-DnaA interaction. In: Biochemical Journal. 2002 ; Vol. 365, No. 3. pp. 881-887.
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AU - Mizushima, Tohru

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