Intracellular autoregulation of the Mycobacterium tuberculosis PrrA response regulator.
Two-component systems are major regulatory systems for bacterial adaptation to environmental changes. During the infectious cycle of Mycobacterium tuberculosis, adaptation to an intracellular environment is critical for multiplication and survival of the micro-organism within the host. The M. tuberculosis prrA gene, encoding the regulator of the two-component system PrrA-PrrB, has been shown to be induced upon macrophage phagocytosis and to be transiently required for the early stages of macrophage infection. In order to study the mechanisms of regulation of the PrrA-PrrB two-component system, PrrA and the cytoplasmic part of the PrrB histidine kinase were produced and purified as hexahistidine-tagged recombinant proteins. Electrophoretic mobility shift assays indicated that PrrA specifically binds to the promoter of its own operon, with increased affinity upon phosphorylation. Moreover, induction of fluorescence was observed after phagocytosis of a wild-type M. tuberculosis strain containing the gfp reporter gene under the control of the prrA-prrB promoter, while this induction was not seen in a prrA/B mutant strain containing the same construct. These results indicate that the early intracellular induction of prrA depends on the autoregulation of this two-component system.
Ewann
Fanny
F
INSERM U447, Institut Pasteur de Lille, 1 rue du Professeur Calmette, F-59019 Lille Cedex, France.
Locht
Camille
C
INSERM U447, Institut Pasteur de Lille, 1 rue du Professeur Calmette, F-59019 Lille Cedex, France.
Supply
Philip
P
INSERM U447, Institut Pasteur de Lille, 1 rue du Professeur Calmette, F-59019 Lille Cedex, France.
D000222:Adaptation, Physiological; D001426:Bacterial Proteins / Q000235:genetics / Q000502:physiology*; D001483:Base Sequence; D004269:DNA, Bacterial / Q000235:genetics / Q000378:metabolism; D005798:Genes, Bacterial; D000071677:Histidine Kinase; D006706:Homeostasis; D009169:Mycobacterium tuberculosis / Q000235:genetics / Q000472:pathogenicity / Q000502:physiology*; D009876:Operon; D011494:Protein Kinases / Q000235:genetics / Q000502:physiology; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D015534:Trans-Activators / Q000235:genetics / Q000502:physiology*
D001426:Bacterial Proteins; D004269:DNA, Bacterial; C085225:REGA protein, Rhodobacter sphaeroides; D011994:Recombinant Proteins; D015534:Trans-Activators; D011494:Protein Kinases; D000071677:Histidine Kinase