Mutational analysis of the histidine operon promoter of Salmonella typhimurium

R. F. Shand, P. H. Blum, D. L. Holzschu, M. S. Urdea, S. W. Artz

Research output: Contribution to journalArticle

5 Citations (Scopus)

Abstract

We isolated a collection of 67 independent, spontaneous Salmonella typhimurium his operon promoter mutants with decreased his expression. The mutants were isolated by selecting for resistance to the toxic lactose analog o-nitrophenyl-β-D-thiogalactoside in a his-lac fusion strain. The collection included base pair substitutions, small insertions, a deletion, and one large insertion identified as IS30 (IS121), which is resident on the Mu d1 cts(Ap(r)lac) phage used to construct the his-lac fusion. Of the 37 mutations that were sequenced, 14 were unique. Six of the 14 were isolated more than once, with the IS30 insertion occurring 16 times. The mutations were located throughout the his promoter region, with two in the conserved -35 hexamer sequence, four in the conserved -10 hexamer sequence (Pribnow box), seven in the spacer between the -10 and -35 hexamer sequences, and the IS30 insertions just upstream of the -35 hexamer sequence. Four of the five substitution mutations changed a consensus base pair recognized by Eσ70 RNA polymerase in the -10 or -35 hexamer. Decreased his expression caused by the 14 different his promoter mutations was measured in vivo. Relative to the wild-type promoter, the mutations resulted in as little as a 4-fold decrease to as much as a 357-fold decrease in his expression, with the larges decreases resulting from changes in the most highly conserved features of Eσ70 promoters.

Original languageEnglish (US)
Pages (from-to)6330-6337
Number of pages8
JournalJournal of bacteriology
Volume171
Issue number11
DOIs
StatePublished - Jan 1 1989

Fingerprint

Salmonella typhimurium
Operon
Histidine
Mutation
Base Pairing
Thiogalactosides
Insertional Mutagenesis
Poisons
DNA-Directed RNA Polymerases
Lactose
Genetic Promoter Regions
Bacteriophages

ASJC Scopus subject areas

  • Microbiology
  • Molecular Biology

Cite this

Mutational analysis of the histidine operon promoter of Salmonella typhimurium. / Shand, R. F.; Blum, P. H.; Holzschu, D. L.; Urdea, M. S.; Artz, S. W.

In: Journal of bacteriology, Vol. 171, No. 11, 01.01.1989, p. 6330-6337.

Research output: Contribution to journalArticle

Shand, R. F. ; Blum, P. H. ; Holzschu, D. L. ; Urdea, M. S. ; Artz, S. W. / Mutational analysis of the histidine operon promoter of Salmonella typhimurium. In: Journal of bacteriology. 1989 ; Vol. 171, No. 11. pp. 6330-6337.
@article{4be8b7eac7fd4bb196384bf38e01db5e,
title = "Mutational analysis of the histidine operon promoter of Salmonella typhimurium",
abstract = "We isolated a collection of 67 independent, spontaneous Salmonella typhimurium his operon promoter mutants with decreased his expression. The mutants were isolated by selecting for resistance to the toxic lactose analog o-nitrophenyl-β-D-thiogalactoside in a his-lac fusion strain. The collection included base pair substitutions, small insertions, a deletion, and one large insertion identified as IS30 (IS121), which is resident on the Mu d1 cts(Ap(r)lac) phage used to construct the his-lac fusion. Of the 37 mutations that were sequenced, 14 were unique. Six of the 14 were isolated more than once, with the IS30 insertion occurring 16 times. The mutations were located throughout the his promoter region, with two in the conserved -35 hexamer sequence, four in the conserved -10 hexamer sequence (Pribnow box), seven in the spacer between the -10 and -35 hexamer sequences, and the IS30 insertions just upstream of the -35 hexamer sequence. Four of the five substitution mutations changed a consensus base pair recognized by Eσ70 RNA polymerase in the -10 or -35 hexamer. Decreased his expression caused by the 14 different his promoter mutations was measured in vivo. Relative to the wild-type promoter, the mutations resulted in as little as a 4-fold decrease to as much as a 357-fold decrease in his expression, with the larges decreases resulting from changes in the most highly conserved features of Eσ70 promoters.",
author = "Shand, {R. F.} and Blum, {P. H.} and Holzschu, {D. L.} and Urdea, {M. S.} and Artz, {S. W.}",
year = "1989",
month = "1",
day = "1",
doi = "10.1128/jb.171.11.6330-6337.1989",
language = "English (US)",
volume = "171",
pages = "6330--6337",
journal = "Journal of Bacteriology",
issn = "0021-9193",
publisher = "American Society for Microbiology",
number = "11",

}

TY - JOUR

T1 - Mutational analysis of the histidine operon promoter of Salmonella typhimurium

AU - Shand, R. F.

AU - Blum, P. H.

AU - Holzschu, D. L.

AU - Urdea, M. S.

AU - Artz, S. W.

PY - 1989/1/1

Y1 - 1989/1/1

N2 - We isolated a collection of 67 independent, spontaneous Salmonella typhimurium his operon promoter mutants with decreased his expression. The mutants were isolated by selecting for resistance to the toxic lactose analog o-nitrophenyl-β-D-thiogalactoside in a his-lac fusion strain. The collection included base pair substitutions, small insertions, a deletion, and one large insertion identified as IS30 (IS121), which is resident on the Mu d1 cts(Ap(r)lac) phage used to construct the his-lac fusion. Of the 37 mutations that were sequenced, 14 were unique. Six of the 14 were isolated more than once, with the IS30 insertion occurring 16 times. The mutations were located throughout the his promoter region, with two in the conserved -35 hexamer sequence, four in the conserved -10 hexamer sequence (Pribnow box), seven in the spacer between the -10 and -35 hexamer sequences, and the IS30 insertions just upstream of the -35 hexamer sequence. Four of the five substitution mutations changed a consensus base pair recognized by Eσ70 RNA polymerase in the -10 or -35 hexamer. Decreased his expression caused by the 14 different his promoter mutations was measured in vivo. Relative to the wild-type promoter, the mutations resulted in as little as a 4-fold decrease to as much as a 357-fold decrease in his expression, with the larges decreases resulting from changes in the most highly conserved features of Eσ70 promoters.

AB - We isolated a collection of 67 independent, spontaneous Salmonella typhimurium his operon promoter mutants with decreased his expression. The mutants were isolated by selecting for resistance to the toxic lactose analog o-nitrophenyl-β-D-thiogalactoside in a his-lac fusion strain. The collection included base pair substitutions, small insertions, a deletion, and one large insertion identified as IS30 (IS121), which is resident on the Mu d1 cts(Ap(r)lac) phage used to construct the his-lac fusion. Of the 37 mutations that were sequenced, 14 were unique. Six of the 14 were isolated more than once, with the IS30 insertion occurring 16 times. The mutations were located throughout the his promoter region, with two in the conserved -35 hexamer sequence, four in the conserved -10 hexamer sequence (Pribnow box), seven in the spacer between the -10 and -35 hexamer sequences, and the IS30 insertions just upstream of the -35 hexamer sequence. Four of the five substitution mutations changed a consensus base pair recognized by Eσ70 RNA polymerase in the -10 or -35 hexamer. Decreased his expression caused by the 14 different his promoter mutations was measured in vivo. Relative to the wild-type promoter, the mutations resulted in as little as a 4-fold decrease to as much as a 357-fold decrease in his expression, with the larges decreases resulting from changes in the most highly conserved features of Eσ70 promoters.

UR - http://www.scopus.com/inward/record.url?scp=0024452244&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0024452244&partnerID=8YFLogxK

U2 - 10.1128/jb.171.11.6330-6337.1989

DO - 10.1128/jb.171.11.6330-6337.1989

M3 - Article

C2 - 2553676

AN - SCOPUS:0024452244

VL - 171

SP - 6330

EP - 6337

JO - Journal of Bacteriology

JF - Journal of Bacteriology

SN - 0021-9193

IS - 11

ER -