Suppression of temperature-sensitivity of a dnaA46 mutant by excessive DNA supercoiling

Taemi Kondo, Shinji Mima, Nobuko Fukuma, Kasuhisa Sekimizu, Tomofusa Tsuchiya, Tohru Mizushima

Research output: Contribution to journalArticle

7 Citations (Scopus)

Abstract

We report here that the high-temperature sensitivity of a dnaA46 mutant was suppressed by addition of high concentrations of NaCl into the culture medium. This suppression was also observed with other high-temperature-sensitive dnaA mutants, except dnaA 167 and dnaA508 mutants, which have mutations in the N-terminal region of DnaA protein. Since high concentrations of NaCl in the medium increased negative DNA supercoiling in a dnaA46 mutant, we hypothesized that the increase in DNA supercoiling is involved in the suppression of the temperature sensitivity of the dnaA46 mutant by high concentrations of NaCl. This hypothesis was supported by in vitro and in vivo results as follows. A low DNA replication activity of purified DnaA46 protein at high temperatures was increased in line with an increase in DNA supercoiling of template DNA. The dnaA46 mutant showed higher sensitivity to nalidixic acid, a DNA-relaxing drug, than did the wild-type cells under the conditions of high temperatures and high concentrations of NaCl.

Original languageEnglish
Pages (from-to)375-379
Number of pages5
JournalBiochemical Journal
Volume348
Issue number2
DOIs
Publication statusPublished - 2000 Jun 1
Externally publishedYes

Fingerprint

Temperature
DNA
Nalidixic Acid
DNA Replication
Culture Media
Proteins
Mutation
Pharmaceutical Preparations

Keywords

  • DNA replication
  • DnaA protein
  • Escherichia coli
  • Linking number
  • Nalidixic acid

ASJC Scopus subject areas

  • Biochemistry

Cite this

Kondo, T., Mima, S., Fukuma, N., Sekimizu, K., Tsuchiya, T., & Mizushima, T. (2000). Suppression of temperature-sensitivity of a dnaA46 mutant by excessive DNA supercoiling. Biochemical Journal, 348(2), 375-379. https://doi.org/10.1042/0264-6021:3480375

Suppression of temperature-sensitivity of a dnaA46 mutant by excessive DNA supercoiling. / Kondo, Taemi; Mima, Shinji; Fukuma, Nobuko; Sekimizu, Kasuhisa; Tsuchiya, Tomofusa; Mizushima, Tohru.

In: Biochemical Journal, Vol. 348, No. 2, 01.06.2000, p. 375-379.

Research output: Contribution to journalArticle

Kondo, T, Mima, S, Fukuma, N, Sekimizu, K, Tsuchiya, T & Mizushima, T 2000, 'Suppression of temperature-sensitivity of a dnaA46 mutant by excessive DNA supercoiling', Biochemical Journal, vol. 348, no. 2, pp. 375-379. https://doi.org/10.1042/0264-6021:3480375
Kondo, Taemi ; Mima, Shinji ; Fukuma, Nobuko ; Sekimizu, Kasuhisa ; Tsuchiya, Tomofusa ; Mizushima, Tohru. / Suppression of temperature-sensitivity of a dnaA46 mutant by excessive DNA supercoiling. In: Biochemical Journal. 2000 ; Vol. 348, No. 2. pp. 375-379.
@article{870e18c60947499ead6894353baa783f,
title = "Suppression of temperature-sensitivity of a dnaA46 mutant by excessive DNA supercoiling",
abstract = "We report here that the high-temperature sensitivity of a dnaA46 mutant was suppressed by addition of high concentrations of NaCl into the culture medium. This suppression was also observed with other high-temperature-sensitive dnaA mutants, except dnaA 167 and dnaA508 mutants, which have mutations in the N-terminal region of DnaA protein. Since high concentrations of NaCl in the medium increased negative DNA supercoiling in a dnaA46 mutant, we hypothesized that the increase in DNA supercoiling is involved in the suppression of the temperature sensitivity of the dnaA46 mutant by high concentrations of NaCl. This hypothesis was supported by in vitro and in vivo results as follows. A low DNA replication activity of purified DnaA46 protein at high temperatures was increased in line with an increase in DNA supercoiling of template DNA. The dnaA46 mutant showed higher sensitivity to nalidixic acid, a DNA-relaxing drug, than did the wild-type cells under the conditions of high temperatures and high concentrations of NaCl.",
keywords = "DNA replication, DnaA protein, Escherichia coli, Linking number, Nalidixic acid",
author = "Taemi Kondo and Shinji Mima and Nobuko Fukuma and Kasuhisa Sekimizu and Tomofusa Tsuchiya and Tohru Mizushima",
year = "2000",
month = "6",
day = "1",
doi = "10.1042/0264-6021:3480375",
language = "English",
volume = "348",
pages = "375--379",
journal = "Biochemical Journal",
issn = "0264-6021",
publisher = "Portland Press Ltd.",
number = "2",

}

TY - JOUR

T1 - Suppression of temperature-sensitivity of a dnaA46 mutant by excessive DNA supercoiling

AU - Kondo, Taemi

AU - Mima, Shinji

AU - Fukuma, Nobuko

AU - Sekimizu, Kasuhisa

AU - Tsuchiya, Tomofusa

AU - Mizushima, Tohru

PY - 2000/6/1

Y1 - 2000/6/1

N2 - We report here that the high-temperature sensitivity of a dnaA46 mutant was suppressed by addition of high concentrations of NaCl into the culture medium. This suppression was also observed with other high-temperature-sensitive dnaA mutants, except dnaA 167 and dnaA508 mutants, which have mutations in the N-terminal region of DnaA protein. Since high concentrations of NaCl in the medium increased negative DNA supercoiling in a dnaA46 mutant, we hypothesized that the increase in DNA supercoiling is involved in the suppression of the temperature sensitivity of the dnaA46 mutant by high concentrations of NaCl. This hypothesis was supported by in vitro and in vivo results as follows. A low DNA replication activity of purified DnaA46 protein at high temperatures was increased in line with an increase in DNA supercoiling of template DNA. The dnaA46 mutant showed higher sensitivity to nalidixic acid, a DNA-relaxing drug, than did the wild-type cells under the conditions of high temperatures and high concentrations of NaCl.

AB - We report here that the high-temperature sensitivity of a dnaA46 mutant was suppressed by addition of high concentrations of NaCl into the culture medium. This suppression was also observed with other high-temperature-sensitive dnaA mutants, except dnaA 167 and dnaA508 mutants, which have mutations in the N-terminal region of DnaA protein. Since high concentrations of NaCl in the medium increased negative DNA supercoiling in a dnaA46 mutant, we hypothesized that the increase in DNA supercoiling is involved in the suppression of the temperature sensitivity of the dnaA46 mutant by high concentrations of NaCl. This hypothesis was supported by in vitro and in vivo results as follows. A low DNA replication activity of purified DnaA46 protein at high temperatures was increased in line with an increase in DNA supercoiling of template DNA. The dnaA46 mutant showed higher sensitivity to nalidixic acid, a DNA-relaxing drug, than did the wild-type cells under the conditions of high temperatures and high concentrations of NaCl.

KW - DNA replication

KW - DnaA protein

KW - Escherichia coli

KW - Linking number

KW - Nalidixic acid

UR - http://www.scopus.com/inward/record.url?scp=0034213190&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0034213190&partnerID=8YFLogxK

U2 - 10.1042/0264-6021:3480375

DO - 10.1042/0264-6021:3480375

M3 - Article

C2 - 10816432

AN - SCOPUS:0034213190

VL - 348

SP - 375

EP - 379

JO - Biochemical Journal

JF - Biochemical Journal

SN - 0264-6021

IS - 2

ER -