Biological and Pharmaceutical Bulletin
Online ISSN : 1347-5215
Print ISSN : 0918-6158
ISSN-L : 0918-6158
Identification of a High-Copy-Number Plasmid Suppressor of a Lethal Phenotype Caused by Mutant DnaA Protein Which Has Decreased Instrinsic ATPase Activity
Masaki MAKISEKenji SAKAMOTOTomofusa TSUCHIYATohru MIZUSHIMA
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1999 年 22 巻 9 号 p. 904-909

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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.

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© The Pharmaceutical Society of Japan
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