ABSTRACT
A number of single-nucleotide polymorphisms (SNPs) have been identified in the genome of Mycobacterium bovis BCG Pasteur compared with the sequenced strain M. bovis 2122/97. The functional consequences of many of these mutations remain to be described; however, mutations in genes encoding regulators may be particularly relevant to global phenotypic changes such as loss of virulence, since alteration of a regulator's function will affect the expression of a wide range of genes. One such SNP falls in bcg3145, encoding a member of the AfsR/DnrI/SARP class of global transcriptional regulators, that replaces a highly conserved glutamic acid residue at position 159 (E159G) with glycine in a tetratricopeptide repeat (TPR) located in the bacterial transcriptional activation (BTA) domain of BCG3145. TPR domains are associated with protein-protein interactions, and a conserved core (helices T1-T7) of the BTA domain seems to be required for proper function of SARP-family proteins. Structural modelling predicted that the E159G mutation perturbs the third alpha-helix of the BTA domain and could therefore have functional consequences. The E159G SNP was found to be present in all BCG strains, but absent from virulent M. bovis and Mycobacterium tuberculosis strains. By overexpressing BCG3145 and Rv3124 in BCG and H37Rv and monitoring transcriptome changes using microarrays, we determined that BCG3145/Rv3124 acts as a positive transcriptional regulator of the molybdopterin biosynthesis moa1 locus, and we suggest that rv3124 be renamed moaR1. The SNP in bcg3145 was found to have a subtle effect on the activity of MoaR1, suggesting that this mutation is not a key event in the attenuation of BCG.
Subject(s)
Coenzymes/biosynthesis , Gene Expression Regulation, Bacterial , Metalloproteins/biosynthesis , Mycobacterium bovis/metabolism , Mycobacterium tuberculosis/metabolism , Polymorphism, Single Nucleotide , Transcription Factors/metabolism , Transcription, Genetic , Amino Acid Motifs , Bacterial Proteins/chemistry , Bacterial Proteins/genetics , Bacterial Proteins/metabolism , Coenzymes/genetics , Metalloproteins/genetics , Molecular Sequence Data , Molybdenum Cofactors , Mycobacterium bovis/chemistry , Mycobacterium bovis/genetics , Mycobacterium tuberculosis/chemistry , Mycobacterium tuberculosis/genetics , Pteridines , Transcription Factors/chemistry , Transcription Factors/geneticsABSTRACT
A novel extremely halophilic archaeon was isolated from Lake Tebenquiche, situated in the northern part of the Atacama Saltern, Chile. The cells of these micro-organisms were mostly irregularly disc-shaped. They grew in medium containing saturated concentrations of NaCl and did not require magnesium for optimal growth. The polar lipid composition revealed the presence of mannosyl-2-sulfate-(1-4)-glycosyl-archaeol, the main glycolipid of the genus Halorubrum, and two new glycolipids. The G+C content of the DNA was 63.2 mol%. Phylogenetic analysis of the 16S rRNA gene placed strain ALT6-92T within the Halorubrum cluster. The low DNA-DNA hybridization value justified classification in a new species for which the name Halorubrum tebenquichense sp. nov. is proposed. The type strain is ALT6-92T (= CECT 5317T = DSM 14210T).
