Abstract
In order to study the regulation of a large block of contiguous genes at the rfa locus of Escherichia coli K-12 which are involved in synthesis and modification of the lipopolysaccharide core, the transposon TnlacZ was used to generate in-frame lacZ fusions to the coding regions of five genes (rfaQ, -G, -P, -B and -J) within this block. The beta-galactosidase activity of strains in which these fusions had been crossed into the chromosomal rfa locus was significantly decreased when the rfaH11 (sfrB11) allele was introduced and was restored to wild-type levels when these strains were lysogenized with a lambda phage carrying wild-type rfaH. This indicates that the positive regulatory function encoded by rfaH is required throughout this block of genes. In addition, expression of the lacZ fusion to rfaJ was reduced by growth at 42 degrees C, and this correlated with a temperature-induced change in the electrophoretic profile of the core lipopolysaccharide.
MeSH Terms
DNA Transposable Elements/genetics
Electrophoresis, Polyacrylamide Gel
Escherichia coli/genetics
Gene Expression Regulation, Bacterial/genetics
Genes, Bacterial
Lipopolysaccharides/genetics
Plasmids/genetics
Recombinant Fusion Proteins/biosynthesis
Temperature
Transcription Factors/genetics
beta-Galactosidase/genetics
Chemicals
DNA Transposable Elements
Lipopolysaccharides
Recombinant Fusion Proteins
Transcription Factors
beta-Galactosidase
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Pradel E
Department of Microbiology, Arizona State University, Tempe 85287-2701.
Schnaitman C A
References (13)
13 references, click to expand
-
Cloning and analysis of the sfrB (sex factor repression) gene of Escherichia coli K-12.
J Bacteriol. 1986 May;166(2):651-7
PMID: 3009418
-
Site-directed insertion and deletion mutagenesis with cloned fragments in Escherichia coli.
J Bacteriol. 1985 Mar;161(3):1219-21
PMID: 2982787
-
Evidence that Rho and NusA are involved in termination in the rplL-rpoB intercistronic region.
J Bacteriol. 1987 May;169(5):2277-80
PMID: 2437101
-
In vitro insertional mutagenesis with a selectable DNA fragment.
Gene. 1984 Sep;29(3):303-13
PMID: 6237955
-
Efficient incorporation of galactose into lipopolysaccharide by Escherichia coli K-12 strains with polar galE mutations.
J Bacteriol. 1990 Sep;172(9):5511-3
PMID: 2168387
-
Genetic analysis of lipopolysaccharide core biosynthesis by Escherichia coli K-12: insertion mutagenesis of the rfa locus.
J Bacteriol. 1990 Sep;172(9):5312-25
PMID: 2168379
-
Two Escherichia coli chromosomal cistrons, sfrA and sfrB, which are needed for expression of F factor tra functions.
J Bacteriol. 1979 Sep;139(3):730-7
PMID: 383690
-
Increased resolution of lipopolysaccharides and lipooligosaccharides utilizing tricine-sodium dodecyl sulfate-polyacrylamide gel electrophoresis.
J Immunol Methods. 1990 Jan 24;126(1):109-17
PMID: 2106001
-
Linkage map of Escherichia coli K-12, edition 8.
Microbiol Rev. 1990 Jun;54(2):130-97
PMID: 2194094
-
Gene rfaH, which affects lipopolysaccharide core structure in Salmonella typhimurium, is required also for expression of F-factor functions.
J Bacteriol. 1981 May;146(2):535-41
PMID: 6111554
-
Regulation of membrane glycosyltransferases by the sfrB and rfaH genes of Escherichia coli and Salmonella typhimurium.
J Biol Chem. 1984 Mar 10;259(5):3064-9
PMID: 6230355
-
sfrA and sfrB products of Escherichia coli K-12 are transcriptional control factors.
J Bacteriol. 1981 Feb;145(2):840-4
PMID: 7007352
-
Analysis of protein localization by use of gene fusions with complementary properties.
J Bacteriol. 1990 Feb;172(2):1035-42
PMID: 2153653