Identification of a high-copy-number plasmid suppressor of a lethal phenotype caused by mutant DnaA protein which has decreased intrinsic ATPase activity

Masaki Makise, Kenji Sakamoto, Tomofusa Tsuchiya, Tohru Mizushima

Research output: Contribution to journalArticle

2 Citations (Scopus)

Abstract

The induction of a mutant DnaA protein (DnaA E204Q) with decreased intrinsic ATPase activity in cells causes a lethal phenotype. Based on our results that the decreased ATPase activity of DnaA E204Q is activated to the level of a wild-type protein in the presence of partially purified stimulating factors for DnaA ATPase, we tested the hypothesis that genes encoding the stimulating factors can be cloned as high-copy-number plasmid suppressors for the lethality caused by DnaA E204Q. We isolated a number of high copy-number plasmids that suppress the lethal phenotype. The genes responsible for the suppression of the lethal phenotype were revealed to be dnaN, relA, pcnB and cyaA by subcloning, and a site-directed mutational analysis. The mechanisms by which these genes suppressed the lethal phenotype were examined in vivo and in vitro.

Original languageEnglish
Pages (from-to)904-909
Number of pages6
JournalBiological and Pharmaceutical Bulletin
Volume22
Issue number9
Publication statusPublished - 1999 Sep
Externally publishedYes

Fingerprint

Mutant Proteins
Adenosine Triphosphatases
Plasmids
Phenotype
Lethal Genes
Genes
Proteins

Keywords

  • ATPase
  • DNA replication
  • DnaA
  • Suppressor
  • The dnaN gene

ASJC Scopus subject areas

  • Molecular Medicine
  • Pharmacology, Toxicology and Pharmaceutics(all)

Cite this

Identification of a high-copy-number plasmid suppressor of a lethal phenotype caused by mutant DnaA protein which has decreased intrinsic ATPase activity. / Makise, Masaki; Sakamoto, Kenji; Tsuchiya, Tomofusa; Mizushima, Tohru.

In: Biological and Pharmaceutical Bulletin, Vol. 22, No. 9, 09.1999, p. 904-909.

Research output: Contribution to journalArticle

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