E-BUGS-93 - Transcription profiling by array of Mycobacterium bovis and Mycobacterium tuberculosis overexpressing overexpressing BCG3145 and Rv3124

Status
Released on 11 October 2010, last updated on 6 August 2014
Organism
Mycobacterium bovis, Mycobacterium tuberculosis
Samples (7)
Array (1)
Protocols (9)
Description
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. Data is also available from http://bugs.sgul.ac.uk/E-BUGS-93
Experiment types
transcription profiling by array, genetic modification, replicate
Contact
Citation
Characterization of the transcriptional regulator Rv3124 of Mycobacterium tuberculosis identifies it as a positive regulator of molybdopterin biosynthesis and defines the functional consequences of a non-synonymous SNP in the Mycobacterium bovis BCG orthologue. Mendoza Lopez P, Golby P, Wooff E, Nunez Garcia J, Garcia Pelayo MC, Conlon K, Gema Camacho A, Hewinson RG, Polaina J, Suarez Garcia A, Gordon SV. Microbiology 156(pt 7):2112-2123 (2010), Europe PMC 20378651
MIAME
PlatformsProtocolsFactorsProcessedRaw
Files
Investigation descriptionE-BUGS-93.idf.txt
Sample and data relationshipE-BUGS-93.sdrf.txt
Raw data (1)E-BUGS-93.raw.1.zip
Processed data (1)E-BUGS-93.processed.1.zip
Experiment designE-BUGS-93.biosamples.png, E-BUGS-93.biosamples.svg
Array designA-BUGS-31.adf.txt
Links