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 language | English |
|---|---|
| Pages (from-to) | 904-909 |
| Number of pages | 6 |
| Journal | Biological and Pharmaceutical Bulletin |
| Volume | 22 |
| Issue number | 9 |
| Publication status | Published - 1999 Sep |
| Externally published | Yes |
Fingerprint
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 journal › Article
}
TY - JOUR
T1 - Identification of a high-copy-number plasmid suppressor of a lethal phenotype caused by mutant DnaA protein which has decreased intrinsic ATPase activity
AU - Makise, Masaki
AU - Sakamoto, Kenji
AU - Tsuchiya, Tomofusa
AU - Mizushima, Tohru
PY - 1999/9
Y1 - 1999/9
N2 - 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.
AB - 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.
KW - ATPase
KW - DNA replication
KW - DnaA
KW - Suppressor
KW - The dnaN gene
UR - http://www.scopus.com/inward/record.url?scp=0032876940&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=0032876940&partnerID=8YFLogxK
M3 - Article
C2 - 10513610
AN - SCOPUS:0032876940
VL - 22
SP - 904
EP - 909
JO - Biological and Pharmaceutical Bulletin
JF - Biological and Pharmaceutical Bulletin
SN - 0918-6158
IS - 9
ER -