Purification and characterization of a repressor for the Bacillus cereus glnRA operon.
We report the overexpression, purification, and properties of the regulatory protein, GlnR, for glutamine synthetase synthesis of Bacillus cereus. The protein was found to be a dimer with a molecular weight of approximately 30,000, and its subunit molecular weight was 15,000 in agreement with that (15,025) of deduced amino acid sequence of GlnR. The purified GlnR protein bound specifically to the promoter region of the glnRA operon of B. cereus and Bacillus subtilis. The binding of the GlnR protein to the DNA fragment was enhanced by the presence of glutamine synthetase, the product of glnA, of B. cereus or B. subtilis, although the affinity of the GlnR protein for DNA was not affected in the presence of glutamate, glutamine, Mg2+, Mn2+, or ammonia. These results indicate the existence of an interaction between GlnR and glutamine synthetase, and support the hypothesis that the regulation of glnA expression requires both GlnR protein and glutamine synthetase in Bacillus.
Nakano
Y
Y
Laboratory of Biochemistry, College of Science, Rikkyo (St. Paul's) University, Tokyo.
Kimura
K
K
D000595:Amino Acid Sequence; D000641:Ammonia / Q000494:pharmacology; D001409:Bacillus cereus / Q000187:drug effects / Q000201:enzymology / Q000235:genetics*; D001483:Base Sequence; D004269:DNA, Bacterial / Q000378:metabolism; D005974:Glutamate-Ammonia Ligase / Q000096:biosynthesis / Q000235:genetics*; D008274:Magnesium / Q000494:pharmacology; D008345:Manganese / Q000494:pharmacology; D008969:Molecular Sequence Data; D008970:Molecular Weight; D009876:Operon; D011401:Promoter Regions, Genetic; D012097:Repressor Proteins / Q000235:genetics* / Q000302:isolation & purification; D015183:Restriction Mapping
D004269:DNA, Bacterial; D012097:Repressor Proteins; D008345:Manganese; D000641:Ammonia; D005974:Glutamate-Ammonia Ligase; D008274:Magnesium