Identification of promoter-binding proteins of the fbp A and C genes in Mycobacterium tuberculosis

Romero, I. C., Mehaffy, C., Burchmore, R. J., Dobos-Elder, K., Brennan, P. and Walker, J. (2010) Identification of promoter-binding proteins of the fbp A and C genes in Mycobacterium tuberculosis. Tuberculosis, 90(1), pp. 25-30. (doi:10.1016/j.tube.2009.10.001)

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Publisher's URL: http://dx.doi.org/10.1016/j.tube.2009.10.001

Abstract

The antigen 85 (Ag85) complex of Mycobacterium tuberculosis represents a promising candidate as a novel drug target and pathogenesis factor. Ag85 comprises three proteins Ag85A, B and C, (encoded by the genes fbpA, B, and C), which participate in cell wall biosynthesis, and interact with the host macrophage as fibronectin-binding proteins (fbps). Ag85 is also involved in the response to isoniazid (INH) treatment. The objective of this study was to identify potential fbp gene activators involved in the over-expression of fbp genes in response to INH. The biotinylated upstream promoter regions of fbpA and fbpC were used together with streptavidin-coated magnetic beads in DNA-binding assays, to isolate proteins with high-binding affinities from cytosolic extracts of INH-treated M. tuberculosis. Resolution of the DNA-binding proteins by 1D SDS-PAGE revealed 6 proteins with high-affinity for the fbpA promoter, and 7 with specificity the fbpC promoter. Mass spectrometric analyses [LC-ES(MS/MS)] identified proteins associated with drug resistance and stress/treatment responses, intermediary metabolism and cellular division, hypothetical proteins including a member of the MarR family of bacterial transcriptional regulators. The DNA-binding MarR protein shows potential as an authentic activator of fbp genes and functional validation of this factor is warranted.

Item Type:Articles
Status:Published
Refereed:Yes
Glasgow Author(s) Enlighten ID:Burchmore, Dr Richard
Authors: Romero, I. C., Mehaffy, C., Burchmore, R. J., Dobos-Elder, K., Brennan, P., and Walker, J.
Subjects:Q Science > Q Science (General)
College/School:College of Medical Veterinary and Life Sciences > Institute of Infection Immunity and Inflammation
Journal Name:Tuberculosis
Publisher:Elsevier
ISSN:1472-9792
ISSN (Online):1873-281X
Published Online:03 December 2009
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Project CodeAward NoProject NamePrincipal InvestigatorFunder's NameFunder RefLead Dept
241291Molecular Genetic and Biochemical Analyses of ParasitesJeremy MottramMedical Research Council (MRC)G97222968Infection Immunity and Inflammation Life Sciences