Identification of amino acids involved in the functional interaction between DnaA protein and acidic phospholipids

Masaki Makise, Shinji Mima, Tomofusa Tsuchiya, Tohru Mizushima

Research output: Contribution to journalArticle

29 Citations (Scopus)

Abstract

DnaA protein, the initiator of chromosomal DNA replication in Escherichia coli, seems to be regulated through its binding to acidic phospholipids, such as cardiolipin. In our previous paper (Hase, M., Yoshimi, T., Ishikawa, Y., Ohba, A., Guo, L., Mima, S., Makise, M., Yamaguchi, Y., Tsuchiya, T., and Mizushima, T. (1998) J. Biol. Chem. 273, 28651-28656), we found that mutant DnaA protein (DnaA431), in which three basic amino acids (Arg360, Arg364, and Lys372) were mutated to acidic amino acids showed a decreased ability to interact with cardiolipin in vitro, suggesting that DnaA protein binds to cardiolipin through an ionic interaction. In this study, we construct three mutant dnaA genes each with a single mutation and examined the function of the mutant proteins in vitro and in vivo. All mutant proteins maintained activities for DNA replication and ATP bind. ing. A mutant protein in which Lys372 was mutated to Glu showed the weakest interaction with cardiolipin among these three mutant proteins. Thus, Lys372 seems to play an important role in the interaction between DnaA protein and acidic phospholipids. Plasmid complementation analyses revealed that all these mutant proteins, including DnaA431 could function as an initiator for chromosomal DNA replication in vivo.

Original languageEnglish
Pages (from-to)4513-4518
Number of pages6
JournalJournal of Biological Chemistry
Volume275
Issue number6
DOIs
Publication statusPublished - 2000 Feb 11
Externally publishedYes

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Mutant Proteins
Cardiolipins
Phospholipids
Amino Acids
DNA Replication
Proteins
DNA
Acidic Amino Acids
Basic Amino Acids
Escherichia coli
Plasmids
Genes
Adenosine Triphosphate
Mutation

ASJC Scopus subject areas

  • Biochemistry

Cite this

Identification of amino acids involved in the functional interaction between DnaA protein and acidic phospholipids. / Makise, Masaki; Mima, Shinji; Tsuchiya, Tomofusa; Mizushima, Tohru.

In: Journal of Biological Chemistry, Vol. 275, No. 6, 11.02.2000, p. 4513-4518.

Research output: Contribution to journalArticle

Makise, Masaki ; Mima, Shinji ; Tsuchiya, Tomofusa ; Mizushima, Tohru. / Identification of amino acids involved in the functional interaction between DnaA protein and acidic phospholipids. In: Journal of Biological Chemistry. 2000 ; Vol. 275, No. 6. pp. 4513-4518.
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