Cloning and characterization of a gene (fadR) involved in regulation of fatty acid metabolism in Escherichia coli

Concetta C DiRusso, W. D. Nunn

Research output: Contribution to journalArticle

27 Citations (Scopus)

Abstract

The regulatory gene fadR has been previously characterized by classical genetic means as a diffusible protein which exerts negative control over fatty acid degradation and acetate metabolism. fadR has also been implicated in the regulation of unsaturated fatty acid biosynthesis. To facilitate the identification of the product of the fadR gene and to study the mechanism by which this multifunctional regulatory gene exerts its control, we cloned a segment of DNA containing the fadR gene in the phage vector λL47. Subsequent subcloning of a segment of the chromosomal DNA from the λfadR+ phage into various plasmid vectors resulted in the isolation of the fadR gene on a 1.3-kilobase-pair HindIII-EcoRV fragment. fadR strains harboring the cloned fadR+ gene showed inducible levels of fatty acid oxidation and crotonase (enoyl-coenzyme A-hydratase, fadB) activity. The cloned gene exerted transcriptional control over β-galactosidase synthesis in an fadR strain that had a λΦ(fadE-lacZ+) operon fusion. An fadR mutation in fabA(Ts) strains prevents growth at permissive temperatures without unsaturated fatty acid supplementation (Nunn et al., J. Bacteriol. 154:554-560, 1983). Plasmids carrying the fadR+ gene suppress this unsaturated fatty acid auxotrophy in fadR fabA(Ts) strains at the permissive condition. Maxicell analysis identified a 29,000-dalton protein encoded by the 1.3-kilobase fragment which appeared to be associated with functional fadR gene activity.

Original languageEnglish (US)
Pages (from-to)583-588
Number of pages6
JournalJournal of bacteriology
Volume161
Issue number2
StatePublished - Jan 1 1985

Fingerprint

Organism Cloning
Fatty Acids
Escherichia coli
Genes
Unsaturated Fatty Acids
Regulator Genes
Bacteriophages
Plasmids
Enoyl-CoA Hydratase
Galactosidases
Hydro-Lyases
DNA
Coenzyme A
Operon
Proteins
Acetates
Mutation
Temperature
Growth

ASJC Scopus subject areas

  • Applied Microbiology and Biotechnology
  • Immunology

Cite this

Cloning and characterization of a gene (fadR) involved in regulation of fatty acid metabolism in Escherichia coli. / DiRusso, Concetta C; Nunn, W. D.

In: Journal of bacteriology, Vol. 161, No. 2, 01.01.1985, p. 583-588.

Research output: Contribution to journalArticle

@article{2f0ee7ee35ae49d4a8185c1f4b21fa4d,
title = "Cloning and characterization of a gene (fadR) involved in regulation of fatty acid metabolism in Escherichia coli",
abstract = "The regulatory gene fadR has been previously characterized by classical genetic means as a diffusible protein which exerts negative control over fatty acid degradation and acetate metabolism. fadR has also been implicated in the regulation of unsaturated fatty acid biosynthesis. To facilitate the identification of the product of the fadR gene and to study the mechanism by which this multifunctional regulatory gene exerts its control, we cloned a segment of DNA containing the fadR gene in the phage vector λL47. Subsequent subcloning of a segment of the chromosomal DNA from the λfadR+ phage into various plasmid vectors resulted in the isolation of the fadR gene on a 1.3-kilobase-pair HindIII-EcoRV fragment. fadR strains harboring the cloned fadR+ gene showed inducible levels of fatty acid oxidation and crotonase (enoyl-coenzyme A-hydratase, fadB) activity. The cloned gene exerted transcriptional control over β-galactosidase synthesis in an fadR strain that had a λΦ(fadE-lacZ+) operon fusion. An fadR mutation in fabA(Ts) strains prevents growth at permissive temperatures without unsaturated fatty acid supplementation (Nunn et al., J. Bacteriol. 154:554-560, 1983). Plasmids carrying the fadR+ gene suppress this unsaturated fatty acid auxotrophy in fadR fabA(Ts) strains at the permissive condition. Maxicell analysis identified a 29,000-dalton protein encoded by the 1.3-kilobase fragment which appeared to be associated with functional fadR gene activity.",
author = "DiRusso, {Concetta C} and Nunn, {W. D.}",
year = "1985",
month = "1",
day = "1",
language = "English (US)",
volume = "161",
pages = "583--588",
journal = "Journal of Bacteriology",
issn = "0021-9193",
publisher = "American Society for Microbiology",
number = "2",

}

TY - JOUR

T1 - Cloning and characterization of a gene (fadR) involved in regulation of fatty acid metabolism in Escherichia coli

AU - DiRusso, Concetta C

AU - Nunn, W. D.

PY - 1985/1/1

Y1 - 1985/1/1

N2 - The regulatory gene fadR has been previously characterized by classical genetic means as a diffusible protein which exerts negative control over fatty acid degradation and acetate metabolism. fadR has also been implicated in the regulation of unsaturated fatty acid biosynthesis. To facilitate the identification of the product of the fadR gene and to study the mechanism by which this multifunctional regulatory gene exerts its control, we cloned a segment of DNA containing the fadR gene in the phage vector λL47. Subsequent subcloning of a segment of the chromosomal DNA from the λfadR+ phage into various plasmid vectors resulted in the isolation of the fadR gene on a 1.3-kilobase-pair HindIII-EcoRV fragment. fadR strains harboring the cloned fadR+ gene showed inducible levels of fatty acid oxidation and crotonase (enoyl-coenzyme A-hydratase, fadB) activity. The cloned gene exerted transcriptional control over β-galactosidase synthesis in an fadR strain that had a λΦ(fadE-lacZ+) operon fusion. An fadR mutation in fabA(Ts) strains prevents growth at permissive temperatures without unsaturated fatty acid supplementation (Nunn et al., J. Bacteriol. 154:554-560, 1983). Plasmids carrying the fadR+ gene suppress this unsaturated fatty acid auxotrophy in fadR fabA(Ts) strains at the permissive condition. Maxicell analysis identified a 29,000-dalton protein encoded by the 1.3-kilobase fragment which appeared to be associated with functional fadR gene activity.

AB - The regulatory gene fadR has been previously characterized by classical genetic means as a diffusible protein which exerts negative control over fatty acid degradation and acetate metabolism. fadR has also been implicated in the regulation of unsaturated fatty acid biosynthesis. To facilitate the identification of the product of the fadR gene and to study the mechanism by which this multifunctional regulatory gene exerts its control, we cloned a segment of DNA containing the fadR gene in the phage vector λL47. Subsequent subcloning of a segment of the chromosomal DNA from the λfadR+ phage into various plasmid vectors resulted in the isolation of the fadR gene on a 1.3-kilobase-pair HindIII-EcoRV fragment. fadR strains harboring the cloned fadR+ gene showed inducible levels of fatty acid oxidation and crotonase (enoyl-coenzyme A-hydratase, fadB) activity. The cloned gene exerted transcriptional control over β-galactosidase synthesis in an fadR strain that had a λΦ(fadE-lacZ+) operon fusion. An fadR mutation in fabA(Ts) strains prevents growth at permissive temperatures without unsaturated fatty acid supplementation (Nunn et al., J. Bacteriol. 154:554-560, 1983). Plasmids carrying the fadR+ gene suppress this unsaturated fatty acid auxotrophy in fadR fabA(Ts) strains at the permissive condition. Maxicell analysis identified a 29,000-dalton protein encoded by the 1.3-kilobase fragment which appeared to be associated with functional fadR gene activity.

UR - http://www.scopus.com/inward/record.url?scp=0021995724&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0021995724&partnerID=8YFLogxK

M3 - Article

VL - 161

SP - 583

EP - 588

JO - Journal of Bacteriology

JF - Journal of Bacteriology

SN - 0021-9193

IS - 2

ER -