Viability and preliminary in vivo characterization of site directed mutants of Escherichia coli single‐stranded DNA‐binding protein

Leslie E. Carlini, Ronald D. Porter, Ute Curth, Claus Urbanke

Research output: Contribution to journalArticle

15 Citations (Scopus)

Abstract

Site‐directed mutations involving selected amino acids of Escherichia coli single‐stranded DNA‐binding protein (SSB) were tested for their in vivo functionality when introduced into a chromosomal ssb deletion strain on a plasmid. All mutants complemented the ssb deletion for viability when present on a pSC101 derivative. The generation time with ssbW54S doubled in comparison to the ssb* control, and both the ssbW54S‐ and ssbH55K‐containing strains exhibited temperature sensitivity. ssbH55K, ssbW54S, ssbW88T, and ssbH55Y ssb‐1 strains displayed reduced survival to ultraviolet irradiation, while ssbW40T and ssbF60L strains were comparable to the ssb+ control strain. This study represents the first investigation of the in vivo properties of ssb mutations constructed for in vitro analysis of DNA binding by SSB.

Original languageEnglish (US)
Pages (from-to)1067-1075
Number of pages9
JournalMolecular Microbiology
Volume10
Issue number5
DOIs
StatePublished - Jan 1 1993

Fingerprint

Escherichia coli
Mutation
Proteins
Plasmids
Amino Acids
Temperature
DNA
In Vitro Techniques

All Science Journal Classification (ASJC) codes

  • Microbiology
  • Molecular Biology

Cite this

Carlini, Leslie E. ; Porter, Ronald D. ; Curth, Ute ; Urbanke, Claus. / Viability and preliminary in vivo characterization of site directed mutants of Escherichia coli single‐stranded DNA‐binding protein. In: Molecular Microbiology. 1993 ; Vol. 10, No. 5. pp. 1067-1075.
@article{6ccac7e6b21040bb86e2c9d6edf598e3,
title = "Viability and preliminary in vivo characterization of site directed mutants of Escherichia coli single‐stranded DNA‐binding protein",
abstract = "Site‐directed mutations involving selected amino acids of Escherichia coli single‐stranded DNA‐binding protein (SSB) were tested for their in vivo functionality when introduced into a chromosomal ssb deletion strain on a plasmid. All mutants complemented the ssb deletion for viability when present on a pSC101 derivative. The generation time with ssbW54S doubled in comparison to the ssb* control, and both the ssbW54S‐ and ssbH55K‐containing strains exhibited temperature sensitivity. ssbH55K, ssbW54S, ssbW88T, and ssbH55Y ssb‐1 strains displayed reduced survival to ultraviolet irradiation, while ssbW40T and ssbF60L strains were comparable to the ssb+ control strain. This study represents the first investigation of the in vivo properties of ssb mutations constructed for in vitro analysis of DNA binding by SSB.",
author = "Carlini, {Leslie E.} and Porter, {Ronald D.} and Ute Curth and Claus Urbanke",
year = "1993",
month = "1",
day = "1",
doi = "10.1111/j.1365-2958.1993.tb00977.x",
language = "English (US)",
volume = "10",
pages = "1067--1075",
journal = "Molecular Microbiology",
issn = "0950-382X",
publisher = "Wiley-Blackwell",
number = "5",

}

Viability and preliminary in vivo characterization of site directed mutants of Escherichia coli single‐stranded DNA‐binding protein. / Carlini, Leslie E.; Porter, Ronald D.; Curth, Ute; Urbanke, Claus.

In: Molecular Microbiology, Vol. 10, No. 5, 01.01.1993, p. 1067-1075.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Viability and preliminary in vivo characterization of site directed mutants of Escherichia coli single‐stranded DNA‐binding protein

AU - Carlini, Leslie E.

AU - Porter, Ronald D.

AU - Curth, Ute

AU - Urbanke, Claus

PY - 1993/1/1

Y1 - 1993/1/1

N2 - Site‐directed mutations involving selected amino acids of Escherichia coli single‐stranded DNA‐binding protein (SSB) were tested for their in vivo functionality when introduced into a chromosomal ssb deletion strain on a plasmid. All mutants complemented the ssb deletion for viability when present on a pSC101 derivative. The generation time with ssbW54S doubled in comparison to the ssb* control, and both the ssbW54S‐ and ssbH55K‐containing strains exhibited temperature sensitivity. ssbH55K, ssbW54S, ssbW88T, and ssbH55Y ssb‐1 strains displayed reduced survival to ultraviolet irradiation, while ssbW40T and ssbF60L strains were comparable to the ssb+ control strain. This study represents the first investigation of the in vivo properties of ssb mutations constructed for in vitro analysis of DNA binding by SSB.

AB - Site‐directed mutations involving selected amino acids of Escherichia coli single‐stranded DNA‐binding protein (SSB) were tested for their in vivo functionality when introduced into a chromosomal ssb deletion strain on a plasmid. All mutants complemented the ssb deletion for viability when present on a pSC101 derivative. The generation time with ssbW54S doubled in comparison to the ssb* control, and both the ssbW54S‐ and ssbH55K‐containing strains exhibited temperature sensitivity. ssbH55K, ssbW54S, ssbW88T, and ssbH55Y ssb‐1 strains displayed reduced survival to ultraviolet irradiation, while ssbW40T and ssbF60L strains were comparable to the ssb+ control strain. This study represents the first investigation of the in vivo properties of ssb mutations constructed for in vitro analysis of DNA binding by SSB.

UR - http://www.scopus.com/inward/record.url?scp=0027739763&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0027739763&partnerID=8YFLogxK

U2 - 10.1111/j.1365-2958.1993.tb00977.x

DO - 10.1111/j.1365-2958.1993.tb00977.x

M3 - Article

VL - 10

SP - 1067

EP - 1075

JO - Molecular Microbiology

JF - Molecular Microbiology

SN - 0950-382X

IS - 5

ER -