Nucleotide Sequence of the Glutamine Synthetase Gene (glnA) and Its Upstream Region from Bacillus cereus

Abstract

We have determined the complete nucleotide sequence of a 2.4 kb chromosomal EcoT22I-NspV fragment, containing the Bacillus cereus glnA gene (structural gene of glutamine synthetase). The deduced amino acid sequence indicates that the glutamine synthetase subunit consists of 444 amino acid residues (50,063 Da). Comparisons are made with reported amino acid sequences of glutamine synthetases from other bacteria. Upstrem of glnA we found an open reading frame of 129 codons (ORF129) preceded by the consensus sequence for a typical promoter. Maxicell experiments showed two polypeptide bands, with molecular weights in good agreement with that of glutamine synthetase and that of ORF129, in addition to vector-coded protein. It is possible that the product of this open reading frame upstream of glnA has a regulatory role in glutamine synthetase expression.

Differential expression of a Clostridium acetobutylicum antisense RNA: implications for regulation of glutamine synthetase

“…Upstream (136 nucleotides) from P1 is such an operator sequence (TGTAAT-5N-ATGAGA). This sequence is very similar to the operator sequences upstream of B. subtilis glnRA and amyR, B. cereus glnA, and E. coli lac and gal genes (TGTNAN-2-5N-NTNACA) (21,25). No convincing operator sequences were located upstream of P3.…”

The Aspergillus nidulans fluG gene is required for production of an extracellular developmental signal and is related to prokaryotic glutamine synthetase I.

“…The most significant matches observed were to glutamine synthetase from the Gram-positive eubacterium Bacillus cereus (28% identity, 50% similarity; Fig. 3) and from the archaebacterium Methanococcus voltae (27% identity, 49% similarity, Nakano et al 1989;Possot et al 1989). Figure 3 shows a BESTFIT comparison between the 436 amino acids at the carboxyl terminus of FluG and B. cereus GlnA.…”

“…In earlier experiments, we constructed a fluG mutant strain by inserting the entire argB gene region into a restriction site in the center of the /iuG-coding se- (Nakano et al 1989). Identities are illustrated by vertical dashes, and conservative changes are illustrated by dots.…”

Purification and characterization of glutamine synthetase from alkalophilic Bacillus No. 170.

“…In contrast, there is no evidence that the Bacillus subtilis glutamine synthetase is regulated by reversible adenylylation although it appears that the mechanisms involved are different from those in E. coli and other enterics. In earlier papers, we reported the isolation and characterization of the glutamine synthetase from both Bacillus cereus and Bacillus subtilis (12,13), the putative amino acid sequence from the DNA sequence of each gene (16,18), and the determination of the substrate binding sites by chemical modification (15,17,24) . To confirm the unique characteristic of the glutamine synthetase from these spore-forming bacteria, we have been investigating nitrogen metabolism in alkalophilic Bacillus.…”

“…Regulation by sulfhydryl group modification in vivo is difficult to establish, however Bacillus glutamine synthetases had a unique, reactive cysteine residue. The modified cysteine residue of B. cereus was Cys 306 (15) that is conserved in Bacillus glutamine synthetase (16,18,22).…”