Transcriptional Regulation of the β-Type Carbonic Anhydrase Gene bca by RamA in Corynebacterium glutamicum

• Published in: PloS one 2016-04, Vol.11 (4), p.e0154382
• Main Authors: Shah, Adnan, Eikmanns, Bernhard J
• Format: Article
• Language: English
• Subjects: Acetates - metabolism; Acetic acid; Alcohol; Amino acids; Bacteria; Bacterial Proteins - genetics; Bacterial Proteins - metabolism; Base Sequence; Bicarbonates; Binding Sites; Biology and Life Sciences; Biotechnology; Carbon dioxide; Carbon sources; Carbonates; Carbonic anhydrase; Carbonic anhydrases; Carbonic Anhydrases - genetics; Carbonic Anhydrases - metabolism; Carboxylation; Clonal deletion; Codon, Initiator; Corynebacterium glutamicum; Corynebacterium glutamicum - genetics; Corynebacterium glutamicum - metabolism; Decarboxylation; Dehydrogenases; Deletion mutant; Enzymes; Gene Deletion; Gene expression; Gene Expression Regulation, Bacterial; Gene regulation; Genes; Glucose - metabolism; Hydration; Kinases; Metabolism; Physical Sciences; Plasmids; Prokaryotes; Promoter Regions, Genetic; Protein Binding; Research and Analysis Methods; Transcription; Transcription Factors - genetics; Transcription Factors - metabolism; Transcription Initiation Site; Transcription, Genetic; Translation
• ISSN: 1932-6203; 1932-6203
• DOI: 10.1371/journal.pone.0154382

Carbonic anhydrase catalyzes the reversible hydration of carbon dioxide to bicarbonate and maintains the balance of CO2/HCO3- in the intracellular environment, specifically for carboxylation/decarboxylation reactions. In Corynebacterium glutamicum, two putative genes, namely the bca (cg2954) and gca (cg0155) genes, coding for β-type and γ-type carbonic anhydrase, respectively, have been identified. We here analyze the transcriptional organization of these genes. The transcriptional start site (TSS) of the bca gene was shown to be the first nucleotide "A" of its putative translational start codon (ATG) and thus, bca codes for a leaderless transcript. The TSS of the gca gene was identified as an "A" residue located at position -20 relative to the first nucleotide of the annotated translational start codon of the cg0154 gene, which is located immediately upstream of gca. Comparative expression analysis revealed carbon source-dependent regulation of the bca gene, with 1.5- to 2-fold lower promoter activity in cells grown on acetate as compared to glucose as sole carbon source. Based on higher expression of bca in a mutant deficient of the regulator of acetate metabolism RamA as compared to the wild-type of C. glutamicum and based on the binding of His-tagged RamA protein to the bca promoter region, we here present evidence that RamA negatively regulates expression of bca in C. glutamicum. Functional characterization of a gca deletion mutant of C. glutamicum revealed the same growth characteristics of C. glutamicum ∆gca as that of wild-type C. glutamicum and no effect on expression of the bca gene.