E-BUGS-91 - Transcription profiling of Mycobacterium tuberculosis using a transposon mutagenesis approach to elucidate pe/ppe regulation

Status
Released on 18 August 2009, last updated on 23 July 2014
Organism
Mycobacterium tuberculosis
Samples (2)
Array (1)
Protocols (7)
Description
The pe/ppe genes are unique to mycobacteria and are widely speculated to play a role in tuberculosis pathogenesis. However, little is known about how expression of these genes is controlled. Elucidating the regulatory control of genes found exclusively in mycobacteria, such as the pe/ppe gene families, may be key to understanding the success of this pathogen. In this study, we used a transposon mutagenesis approach to elucidate pe/ppe regulation. This resulted in the identification of Rv0485, a previously uncharacterized transcriptional regulator. Microarray and quantitative real-time PCR analysis confirmed that disruption of Rv0485 reduced the expression of the pe13/ppe18 gene pair (Rv1195/Rv1196), defined the Rv0485 regulon, and emphasized the lack of global regulation of pe/ppe genes. The in vivo phenotype of the Rv0485 transposon mutant strain (Rv0485::Tn) was investigated in the mouse model, where it was demonstrated that the mutation has minimal effect on bacterial organ burden. Despite this, disruption of Rv0485 allowed mice to survive for significantly longer, with substantially reduced lung pathology in comparison with mice infected with wild type M. tuberculosis. Infection of immune-deficient SCID mice with the Rv0485::Tn strain also resulted in extended survival times, suggesting that Rv0485 plays a role in modulation of innate immune responses. This is further supported by the finding that disruption of Rv0485 resulted in reduced secretion of pro-inflammatory cytokines by infected murine macrophages. In summary, we have demonstrated that disruption of a previously uncharacterized transcriptional regulator, Rv0485, results in reduced expression of pe13/ppe18 and attenuation of M. tuberculosis virulence. Data is also available from http://bugs.sgul.ac.uk/E-BUGS-91
Experiment types
transcription profiling by array, dye swap, strain or line
Contact
Citation
MIAME
PlatformsProtocolsVariablesProcessedRaw
Files
Investigation descriptionE-BUGS-91.idf.txt
Sample and data relationshipE-BUGS-91.sdrf.txt
Raw data (1)E-BUGS-91.raw.1.zip
Processed data (1)E-BUGS-91.processed.1.zip
Experiment designE-BUGS-91.biosamples.png, E-BUGS-91.biosamples.svg
Array designA-BUGS-23.adf.txt
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