Sequence and linkage analysis of the Coxiella burnetii citrate synthase-encoding gene

• Published in: Gene Vol. 109; no. 1; pp. 63 - 69
• Main Authors: Heinzen, Robert A., Frazier, Marvin E., Mallavia, Louis P.
• Format: Journal Article
• Language: English
• Published: Lausanne Elsevier B.V 20.12.1991; Amsterdam Elsevier; New York, NY
• Subjects: Amino Acid Sequence; Base Sequence; Biological and medical sciences; cDNA; Citrate (si)-Synthase - genetics; citrate synthase; Cloning, Molecular; Codon; Coxiella burnetii; Coxiella burnetii - genetics; Fundamental and applied biological sciences. Psychology; genes; Genes. Genome; Genetic Linkage; homology; Molecular and cellular biology; Molecular genetics; Molecular Sequence Data; nucleotide sequence; predictions; Promoter Regions, Genetic; Reading Frames; Recombinant DNA; Repetitive Sequences, Nucleic Acid; rickettsia; Sequence Homology, Nucleic Acid; succinate dehydrogenase; tricarboxylic acid cycle; aa; ExoIII; rickettsia; ORF; RBS; SSC; homology; nt; IR; Recombinant DNA; SdhA; bp; succinate dehydrogenase; dITP; tsp; tricarboxylic acid cycle; GltA; TCA; kb; oligo; REP; Genetic mapping; Linkage; Nucleotide sequence; Enzyme; Homology; Rickettsiales; Citrate (si)-synthase; Bacteria; Chromosome DNA; Rickettsieae; Molecular cloning; Coxiella burnetii; Rickettsiaceae
• ISSN: 0378-1119; 1879-0038
• DOI: 10.1016/0378-1119(91)90589-4

The nucleotide (nt) sequence of the Coxiella burnetii citrate synthase-encoding gene ( gltA), previously cloned in Escherichia coli, was determined. The nt sequence analysis revealed an open reading frame (ORF) of 1290 bp capable of coding for a protein of 430 amino acids (aa) with a deduced M r of 48 633. Preceding an ATG start codon, a possible transcription start point ( tsp) with homology to the E. coli promoter consensus was detected. A poly-purine-rich region occurred immediately upstream from the gltA reading frame and potentially serves as a ribosome-binding site. Additionally, a G + C-rich region of dyad symmetry 3′ to the translational stop codon was found that could possibly function as a Rho-independent transcriptional termination signal. A large, nearly perfect, inverted repeat was identified upstream from the gltA tsp and was shown by Southern analysis to be present in multiple copies in the C. burnetii genome. The deduced aa sequence of C. burnetii GltA was optimally aligned with enzymes from various prokaryotic sources and one eukaryotic source (pig heart). Using perfect aa identity, the C. burnetii enzyme demonstrated the greatest homology with GltA from Acinetobacter anitratum (65%). Although only 26% aa identity was seen with the pig heart enzyme, many of the residues identified in ligand binding appear to be conserved. Sequencing studies of a region centered approx. 5.6 kb upstream from gltA revealed an ORF read with opposite polarity that encodes a peptide highly homologous to the C terminus of the flavoprotein subunit of E. coli succinate dehydrogenase. This report represents the first nt sequence analysis of a gene of known function from the obligate intracellular parasite, C. burnetii.