Cholesterol utilization in mycobacteria is controlled by two TetR-type transcriptional regulators: kstR and kstR2

• DOI: 10.1099/mic.0.034538-0
• Publication Date: 2010-02-19T01:45:19Z
• Authors: Sharon L. Kendall; Philippa Burgess; Ricardo Balhana; Mike Withers; Annemieke ten Bokum; J. Shaun Lott; Chen Gao; Iria Uhia-Castro; Neil G. Stoker
• Keywords: 0301 basic medicine; Binding Sites; Reverse Transcriptase Polymerase Chain Reaction; Amino Acid Motifs; Inverted Repeat Sequences; Mycobacterium smegmatis; Mycobacterium tuberculosis; Regulon; Mycobacterium; 3. Good health; Repressor Proteins; 03 medical and health sciences; Cholesterol; Bacterial Proteins; Gene Expression Regulation; Species Specificity; Cell and Molecular Biology of Microbes; Promoter Regions, Genetic; Conserved Sequence; Oligonucleotide Array Sequence Analysis

Mycobacterium tuberculosis is able to use a variety of carbon sources in vivo and current knowledge suggests that cholesterol is used as a carbon source during infection. The catabolized cholesterol is used both as an energy source (ATP generation) and as a source of precursor molecules for the synthesis of complex methyl-branched fatty acids. In previous studies, we described a TetR-type transcriptional repressor, kstR, that controls the expression of a number of genes involved in cholesterol catabolism. In this study, we describe a second TetR-type repressor, which we call kstR2. We knocked this gene out in Mycobacterium smegmatis and used microarrays and quantitative RT-PCR to examine the effects on gene expression. We identified a palindromic regulatory motif for KstR2, showed that this motif is present in three promoter regions in mycobacteria and rhodococcus, and demonstrated binding of purified KstR2 to the motif. Using a combination of motif location analysis, gene expression analysis and the examination of gene conservation, we suggest that kstR2 controls the expression of a 15 gene regulon. Like kstR, kstR2 and the kstR2 regulon are highly conserved among the actinomycetes and studies in rhodococcus suggest a role for these genes in cholesterol catabolism. The functional significance of the regulon and implications for the control of cholesterol utilization are discussed.