Control of transcription of gal repressor and isorepressor genes in Escherichia coli

Two regulatory proteins, Gal repressor and isorepressor, control the expression of the gal and mgl operons in Escherichia coli. The transcription start sites for galR and galS, the genes for the repressor and isorepressor, were determined by primer extension of in vivo transcripts. Study of the promoter-lacZ gene fusions introduced into the chromosome indicated that galS expression was elevated in cells in which the normal galS gene was interrupted, but not in cells in which the galR gene was deleted. When both genes were disrupted, galS expression was further elevated. Expression from the galS promoter was stimulated by the addition of D-fucose, repressed by glucose, and dependent on cyclic AMP receptor protein (CRP). Expression of a similar gene fusion of the galR promoter to lacZ was unregulated. Both galR and galS genes contain two potential operator sites (OE and OI) and a CRP-binding site. The arrangement of OE, OI, and the CRP-binding site in the galS gene is analogous to the arrangement in the gal and mgl promoters, but the arrangement in galR is atypical. The increased concentration of the isorepressor when inducer is present may facilitate early shutoff of the isorepressor-regulated genes of the gal regulon when inducer (substrate) concentration falls.

[1]  P. D. de Boer,et al.  Isolation and properties of minB, a complex genetic locus involved in correct placement of the division site in Escherichia coli , 1988, Journal of bacteriology.

[2]  L. Enquist,et al.  Experiments With Gene Fusions , 1984 .

[3]  G. Buttin Mécanismes régulateurs dans la biosynthèse des enzymes du métabolisme du galactose chez Escherichia coli K12: I. La biosynthèse induite de la galactokinase et l'induction simultanée de la séquence enzymatique , 1963 .

[4]  H. Kalckar,et al.  HEREDITARY DEFECTS IN GALACTOSE METABOLISM IN ESCHERICHIA COLI MUTANTS, I. DETERMINATION OF ENZYME ACTIVITIES. , 1959, Proceedings of the National Academy of Sciences of the United States of America.

[5]  S. Adhya,et al.  Isorepressor of the gal regulon in Escherichia coli. , 1992, Journal of molecular biology.

[6]  B. Rotman,et al.  Transport systems for galactose and galactosides in Escherichia coli. II. Substrate and inducer specificities. , 1968, Journal of molecular biology.

[7]  J. Shapiro,et al.  The galactose operon of E. coli K-12. II. A deletion analysis of operon structure and polarity. , 1969, Genetics.

[8]  S. Douthwaite,et al.  Design of camp–CRP‐activated promoters in Escherichia coli , 1991, Molecular microbiology.

[9]  S. Adhya,et al.  A mutation defining ultrainduction of the Escherichia coli gal operon , 1991, Journal of bacteriology.

[10]  P. Starlinger,et al.  Negative control of the galactose operon in E. coli , 1968, Molecular and General Genetics MGG.

[11]  S. Adhya,et al.  Cyclic-AMP-dependent switch in initiation of transcription from the two promoters of the Escherichia coli gal operon: identification and assay of 5'-triphosphate ends of mRNA by GTP:RNA guanyltransferase , 1989, Journal of bacteriology.

[12]  Jeffrey H. Miller Experiments in molecular genetics , 1972 .

[13]  A. Travers,et al.  Conserved features of coordinately regulated E. coli promoters. , 1984, Nucleic acids research.

[14]  S. Adhya,et al.  Modulation of the two promoters of the galactose operon of Escherichia coli , 1979, Nature.

[15]  J. Sambrook,et al.  Molecular Cloning: A Laboratory Manual , 2001 .

[16]  J. Chirgwin,et al.  Isolation of RNA. , 1989, Methods in enzymology.

[17]  B. Rotman,et al.  Transport systems for galactose and galactosides in Escherichia coli. I. Genetic determination and regulation of the methyl-galactoside permease. , 1966, Journal of molecular biology.

[18]  P. Nygaard,et al.  Autoregulation of PurR repressor synthesis and involvement of purR in the regulation of purB, purC, purL, purMN and guaBA expression in Escherichia coli. , 1990, European journal of biochemistry.

[19]  P. Valentin‐Hansen,et al.  Transcriptional regulation of the cytR repressor gene of Escherichia coli: autoregulation and positive control by the cAMP/CAP complex , 1990, Molecular Microbiology.

[20]  S. Adhya,et al.  Probing the structure of gal operator-repressor complexes. Conformation change in DNA. , 1987, The Journal of biological chemistry.

[21]  S. Adhya,et al.  A family of bacterial regulators homologous to Gal and Lac repressors. , 1992, The Journal of biological chemistry.

[22]  J. Shapiro,et al.  The galactose operon of E. coli K-12. I. Structural and pleiotropic mutations of the operon. , 1969, Genetics.

[23]  S. Adhya,et al.  Further inducibility of a constitutive system: ultrainduction of the gal operon , 1991, Journal of bacteriology.

[24]  B. Müller-Hill,et al.  Sequence of galR gene indicates a common evolutionary origin of lac and gal repressor in Escherichia coli. , 1982, Proceedings of the National Academy of Sciences of the United States of America.

[25]  A. Travers,et al.  Promoter Sequence for Stringent Control of Bacterial Ribonucleic Acid Synthesis , 1980, Journal of bacteriology.

[26]  S. Adhya,et al.  Demonstration of two operator elements in gal: in vitro repressor binding studies. , 1984, Proceedings of the National Academy of Sciences of the United States of America.

[27]  D. K. Hawley,et al.  Compilation and analysis of Escherichia coli promoter DNA sequences. , 1983, Nucleic acids research.

[28]  R. Rolfes,et al.  Autoregulation of Escherichia coli purR requires two control sites downstream of the promoter , 1990, Journal of bacteriology.