Abstract
Cells of Escherichia coli undergo a metabolic switch associated with the production and utilization of acetate. During exponential growth on tryptone broth, these cells excrete acetate via the phosphotransacetylase-acetate kinase (Pta-AckA) pathway. As they begin the transition to stationary phase, they instead resorb acetate, activate it to acetyl coenzyme A (acetyl-CoA) by means of the enzyme acetyl-CoA synthetase (Acs) and utilize it to generate energy and biosynthetic components via the tricarboxylic acid cycle and the glyoxylate shunt, respectively. This metabolic switch depends upon the induction of Acs. As part of our effort to dissect the mechanism(s) underlying induction and to identify the signal(s) that triggers that induction, we sought the sigma factor most responsible for acs expression. Using isogenic strains that carry a temperature sensitivity allele of the gene that encodes sigma(70) and either a wild-type or null allele of the gene that encodes sigma(S), we determined by immunoblotting, reverse transcriptase PCR, and acs::lacZ transcriptional fusion analyses that sigma(70) is the sigma factor primarily responsible for the acs transcription that cells induce during mid-exponential phase. In contrast, sigma(S) partially inhibits that transcription as cells enter stationary phase.
MeSH Terms
Acetate-CoA Ligase/genetics
Animals
Bacterial Proteins/physiology
DNA-Binding Proteins/physiology
DNA-Directed RNA Polymerases/physiology
Escherichia coli/enzymology,genetics
Polymerase Chain Reaction
Rabbits
Sigma Factor/physiology
Temperature
Transcription, Genetic
beta-Galactosidase/metabolism
Chemicals
Bacterial Proteins
DNA-Binding Proteins
Sigma Factor
RNA polymerase sigma 70
DNA-Directed RNA Polymerases
beta-Galactosidase
Acetate-CoA Ligase
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Kumari S
Department of Microbiology, Stritch School of Medicine, Loyola University Chicago, Maywood, Illinois 60153, USA.
Simel E J
Wolfe A J
References (18)
18 references, click to expand
-
Cleavage of structural proteins during the assembly of the head of bacteriophage T4.
Nature. 1970 Aug 15;227(5259):680-5
PMID: 5432063
-
Heterogeneity of the principal sigma factor in Escherichia coli: the rpoS gene product, sigma 38, is a second principal sigma factor of RNA polymerase in stationary-phase Escherichia coli.
Proc Natl Acad Sci U S A. 1993 Apr 15;90(8):3511-5
PMID: 8475100
-
Regulation of transcription of katE and katF in Escherichia coli.
J Bacteriol. 1990 Dec;172(12):6713-20
PMID: 2254248
-
Involvement of iclR and rpoS in the induction of acs, the gene for acetyl coenzyme A synthetase of Escherichia coli K-12.
FEMS Microbiol Lett. 1997 Jan 1;146(1):103-8
PMID: 8997713
-
Improved single and multicopy lac-based cloning vectors for protein and operon fusions.
Gene. 1987;53(1):85-96
PMID: 3596251
-
Regulation of RNA polymerase sigma subunit synthesis in Escherichia coli: intracellular levels of sigma 70 and sigma 38.
J Bacteriol. 1995 Dec;177(23):6832-5
PMID: 7592475
-
Cloning, characterization, and functional expression of acs, the gene which encodes acetyl coenzyme A synthetase in Escherichia coli.
J Bacteriol. 1995 May;177(10):2878-86
PMID: 7751300
-
Regulation of acetyl phosphate synthesis and degradation, and the control of flagellar expression in Escherichia coli.
Mol Microbiol. 1994 Jun;12(6):973-84
PMID: 7934904
-
Stationary-phase-inducible "gearbox" promoters: differential effects of katF mutations and role of sigma 70.
J Bacteriol. 1991 Jul;173(14):4482-92
PMID: 1906064
-
Negative regulation by RpoS: a case of sigma factor competition.
Mol Microbiol. 1998 Aug;29(4):1039-51
PMID: 9767572
-
Expression of Escherichia coli pyruvate oxidase (PoxB) depends on the sigma factor encoded by the rpoS(katF) gene.
Mol Microbiol. 1994 Mar;11(6):1019-28
PMID: 8022274
-
Mutations in NADH:ubiquinone oxidoreductase of Escherichia coli affect growth on mixed amino acids.
J Bacteriol. 1994 Apr;176(8):2143-50
PMID: 8157582
-
Molecular characterization of the promoter of osmY, an rpoS-dependent gene.
J Bacteriol. 1994 Jan;176(1):100-7
PMID: 8282684
-
Enzymatic phosphorylation of acetate.
J Biol Chem. 1954 Dec;211(2):737-56
PMID: 13221579
-
Control of carbon flux to acetate excretion during growth of Escherichia coli in batch and continuous cultures.
J Gen Microbiol. 1989 Nov;135(11):2875-83
PMID: 2693588
-
[Enzymatic determination of acetate].
Biochem Z. 1966 Mar 28;344(2):167-89
PMID: 5982377
-
Acyl adenylates; an enzymatic mechanism of acetate activation.
J Biol Chem. 1956 Oct;222(2):991-1013
PMID: 13367067
-
A procedure for isolation of spontaneous mutants with temperature sensitive of RNA and/or protein.
Mol Gen Genet. 1977 Nov 18;156(3):233-7
PMID: 340905