1887

Abstract

Two-component systems are major regulatory systems for bacterial adaptation to environmental changes. During the infectious cycle of , adaptation to an intracellular environment is critical for multiplication and survival of the micro-organism within the host. The 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 strain containing the reporter gene under the control of the promoter, while this induction was not seen in a / mutant strain containing the same construct. These results indicate that the early intracellular induction of depends on the autoregulation of this two-component system.

Keyword(s): His6, hexahistidine
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2004-01-01
2020-04-07
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