Isorepressor of the gal regulon in Escherichia coli.

Inducible overexpression of the Escherichia coli gal operon in the absence of the Gal repressor is known as ultrainduction. The requirement of induction can be eliminated by mutation of a new locus, galS, resulting in constitutive and ultrainduced levels of gal expression. Characterization of the galS gene and its product has revealed an isorepressor of the gal regulon. The Gal isorepressor is a protein of 346 amino acid residues whose amino acid sequence and cellular function, as described here, are very similar to that of Gal repressor, encoded by the galR gene. Transcription from different promoters of the gal regulon, galP1, galP2 and mglP, was examined by primer extension and reverse transcription of mRNA isolated from strains containing mutations in galR and/or galS. In strains containing a galS mutation, overexpression of gal message occurred only in the presence of inducer, while mgl message was constitutively derepressed. The galS mutation also constitutively derepressed an mglA::lacZ fusion, demonstrating that GalS is the mgl repressor. A potential operator site in the mgl promoter was identified at a position analogous to OE in gal. Thus, the gal and mgl operons constitute a regulon. Crosstalk, temporal action, induction spectrum or heteromer formation between repressor and isorepressor may help co-ordinate high affinity galactose transport and galactose utilization.

[1]  D. Court,et al.  Isolation of plaque-forming, galactose-transducing strains of phage lambda. , 1972, Genetics.

[2]  J H Miller,et al.  Genetic studies of the lac repressor. XIII. Extensive amino acid replacements generated by the use of natural and synthetic nonsense suppressors. , 1990, Journal of molecular biology.

[3]  D. Lipman,et al.  Improved tools for biological sequence comparison. , 1988, Proceedings of the National Academy of Sciences of the United States of America.

[4]  S. Adhya,et al.  Interaction of spatially separated protein-DNA complexes for control of gene expression: operator conversions. , 1988, Proceedings of the National Academy of Sciences of the United States of America.

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

[6]  J. Lengeler,et al.  The regulation of the beta-methylgalactoside transport system and of the galactose binding protein of Escherichia coli K12. , 1971, European journal of biochemistry.

[7]  G. Cox,et al.  Phosphate-specific transport system of Escherichia coli: nucleotide sequence and gene-polypeptide relationships , 1985, Journal of bacteriology.

[8]  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.

[9]  Locations and orientations on the Escherichia coli physical map of the mgl operon and galS, a new locus for galactose ultrainduction , 1991, Journal of bacteriology.

[10]  M. Hermodson,et al.  The nucleotide sequences of the rbsD, rbsA, and rbsC genes of Escherichia coli K12. , 1986, The Journal of biological chemistry.

[11]  D. Wilson The regulation and properties of the galactose transport system in Escherichia coli K12. , 1974, The Journal of biological chemistry.

[12]  S. Busby,et al.  Cyclic AMP-dependent constitutive expression of gal operon: use of repressor titration to isolate operator mutations. , 1983, Proceedings of the National Academy of Sciences of the United States of America.

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

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

[15]  F. Studier,et al.  Use of bacteriophage T7 RNA polymerase to direct selective high-level expression of cloned genes. , 1986, Journal of molecular biology.

[16]  D. Glover DNA cloning : a practical approach , 1985 .

[17]  F A Quiocho,et al.  Comparison of the periplasmic receptors for L-arabinose, D-glucose/D-galactose, and D-ribose. Structural and Functional Similarity. , 1991, The Journal of biological chemistry.

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

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

[20]  J. Way,et al.  New Tn10 derivatives for transposon mutagenesis and for construction of lacZ operon fusions by transposition. , 1984, Gene.

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

[22]  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.

[23]  S. Rudikoff,et al.  Purification and properties of Gal repressor:pL-galR fusion in pKC31 plasmid vector. , 1987, The Journal of biological chemistry.

[24]  H. C. Wu,et al.  Role of the galactose transport system in the establishment of endogenous induction of the galactose operon in Escherichia coli. , 1967, Journal of molecular biology.

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

[26]  K. Isono,et al.  The physical map of the whole E. coli chromosome: Application of a new strategy for rapid analysis and sorting of a large genomic library , 1987, Cell.

[27]  J. Adler,et al.  Isolation and Complementation of Mutants in Galactose Taxis and Transport , 1974, Journal of bacteriology.

[28]  Characterization of mutations in oligomerization domain of Lac repressor protein. , 1991, The Journal of biological chemistry.

[29]  R. Gennis,et al.  The sequence of the cyo operon indicates substantial structural similarities between the cytochrome o ubiquinol oxidase of Escherichia coli and the aa3-type family of cytochrome c oxidases. , 1990, The Journal of biological chemistry.

[30]  B. Matthews,et al.  The helix-turn-helix DNA binding motif. , 1989, The Journal of biological chemistry.

[31]  R. Kadner,et al.  Nucleotide sequence of the uhp region of Escherichia coli , 1987, Journal of bacteriology.

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

[33]  S. Harrison,et al.  DNA recognition by proteins with the helix-turn-helix motif. , 1990, Annual review of biochemistry.

[34]  U. K. Laemmli,et al.  Cleavage of Structural Proteins during the Assembly of the Head of Bacteriophage T4 , 1970, Nature.

[35]  A. R. Robbins Regulation of the Escherichia coli methylgalactoside transport system by gene mglD , 1975, Journal of bacteriology.

[36]  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.

[37]  W. Szybalski,et al.  Orientation of transcription for the galactose operon as determined by hybridization of gal mRNA with the separated DNA strands of coliphages lambda-dg. , 1968, Journal of molecular biology.

[38]  G. Buttin Mécanismes régulateurs dans la biosynthèse des enzymes du métabolisme du galactose chez Escherichia coli K12: II. Le Déterminisme génétique de la régulation* , 1963 .

[39]  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 .