Isolation and characterization of inhibitory factors of DNA polymerase III holoenzyme from Escherichia coli

Masakazu Hase, Tohru Mizushima, Tsutomu Katayama, Kazuhisa Sekimizu

Research output: Contribution to journalArticle

Abstract

We isolated fractions by Mono Q chromatography that inhibited the activity of Escherichia coli DNA polymerase III holoenzyme using an assay system with a primed single-stranded DNA template coated with single-stranded DNA binding protein (SSB). The inhibitory activities were inactivated by heat-treatment at 100 °C for 10 min, suggesting that they are proteins. The factors did not inhibit the activity of RNA polymerase of Escherichia coli. The inhibitory effects were less potent for the activities of the large (Klenow) fragment of DNA polymerase I and T4 DNA polymerase than for DNA polymerase HI holoenzyme. No degradation of single-or double-stranded DNA was observed in the fractions, indicating that inhibition was not due to degradation of the DNA.

Original languageEnglish
Pages (from-to)215-220
Number of pages6
JournalFEMS microbiology letters
Volume130
Issue number2-3
DOIs
Publication statusPublished - Aug 1 1995

Fingerprint

DNA Polymerase III
DNA Polymerase I
Holoenzymes
DNA-Directed DNA Polymerase
Escherichia coli
Single-Stranded DNA
DNA
DNA-Binding Proteins
DNA-Directed RNA Polymerases
Chromatography
Hot Temperature
Proteins
Mono Q

All Science Journal Classification (ASJC) codes

  • Microbiology
  • Molecular Biology
  • Genetics

Cite this

Isolation and characterization of inhibitory factors of DNA polymerase III holoenzyme from Escherichia coli. / Hase, Masakazu; Mizushima, Tohru; Katayama, Tsutomu; Sekimizu, Kazuhisa.

In: FEMS microbiology letters, Vol. 130, No. 2-3, 01.08.1995, p. 215-220.

Research output: Contribution to journalArticle

Hase, Masakazu ; Mizushima, Tohru ; Katayama, Tsutomu ; Sekimizu, Kazuhisa. / Isolation and characterization of inhibitory factors of DNA polymerase III holoenzyme from Escherichia coli. In: FEMS microbiology letters. 1995 ; Vol. 130, No. 2-3. pp. 215-220.
@article{ae2d00a02bab4de6be2ff53bd6c8f090,
title = "Isolation and characterization of inhibitory factors of DNA polymerase III holoenzyme from Escherichia coli",
abstract = "We isolated fractions by Mono Q chromatography that inhibited the activity of Escherichia coli DNA polymerase III holoenzyme using an assay system with a primed single-stranded DNA template coated with single-stranded DNA binding protein (SSB). The inhibitory activities were inactivated by heat-treatment at 100 °C for 10 min, suggesting that they are proteins. The factors did not inhibit the activity of RNA polymerase of Escherichia coli. The inhibitory effects were less potent for the activities of the large (Klenow) fragment of DNA polymerase I and T4 DNA polymerase than for DNA polymerase HI holoenzyme. No degradation of single-or double-stranded DNA was observed in the fractions, indicating that inhibition was not due to degradation of the DNA.",
author = "Masakazu Hase and Tohru Mizushima and Tsutomu Katayama and Kazuhisa Sekimizu",
year = "1995",
month = "8",
day = "1",
doi = "10.1016/0378-1097(95)00209-N",
language = "English",
volume = "130",
pages = "215--220",
journal = "FEMS Microbiology Letters",
issn = "0378-1097",
publisher = "Wiley-Blackwell",
number = "2-3",

}

TY - JOUR

T1 - Isolation and characterization of inhibitory factors of DNA polymerase III holoenzyme from Escherichia coli

AU - Hase, Masakazu

AU - Mizushima, Tohru

AU - Katayama, Tsutomu

AU - Sekimizu, Kazuhisa

PY - 1995/8/1

Y1 - 1995/8/1

N2 - We isolated fractions by Mono Q chromatography that inhibited the activity of Escherichia coli DNA polymerase III holoenzyme using an assay system with a primed single-stranded DNA template coated with single-stranded DNA binding protein (SSB). The inhibitory activities were inactivated by heat-treatment at 100 °C for 10 min, suggesting that they are proteins. The factors did not inhibit the activity of RNA polymerase of Escherichia coli. The inhibitory effects were less potent for the activities of the large (Klenow) fragment of DNA polymerase I and T4 DNA polymerase than for DNA polymerase HI holoenzyme. No degradation of single-or double-stranded DNA was observed in the fractions, indicating that inhibition was not due to degradation of the DNA.

AB - We isolated fractions by Mono Q chromatography that inhibited the activity of Escherichia coli DNA polymerase III holoenzyme using an assay system with a primed single-stranded DNA template coated with single-stranded DNA binding protein (SSB). The inhibitory activities were inactivated by heat-treatment at 100 °C for 10 min, suggesting that they are proteins. The factors did not inhibit the activity of RNA polymerase of Escherichia coli. The inhibitory effects were less potent for the activities of the large (Klenow) fragment of DNA polymerase I and T4 DNA polymerase than for DNA polymerase HI holoenzyme. No degradation of single-or double-stranded DNA was observed in the fractions, indicating that inhibition was not due to degradation of the DNA.

UR - http://www.scopus.com/inward/record.url?scp=0029100278&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0029100278&partnerID=8YFLogxK

U2 - 10.1016/0378-1097(95)00209-N

DO - 10.1016/0378-1097(95)00209-N

M3 - Article

VL - 130

SP - 215

EP - 220

JO - FEMS Microbiology Letters

JF - FEMS Microbiology Letters

SN - 0378-1097

IS - 2-3

ER -