1887

Abstract

HtrA is a bifunctional stress protein required by many bacterial pathogens to successfully cause infection. serovar Typhimurium ( Typhimurium) mutants are defective in intramacrophage survival and are highly attenuated in mice. Transcription of in is governed by a single promoter that is dependent on (RpoE). Typhimurium also possesses a -dependent promoter; however, we found that the absence of had little effect on production of HtrA by Typhimurium. This suggests that additional promoters control expression of in Typhimurium. We identified three Typhimurium promoters. Only the most proximal promoter, , was dependent. The other promoters, and , are probably recognized by the principal sigma factor . These two promoters were constitutively expressed but were also slightly induced by heat shock. Thus expression of is different in Typhimurium and . The role of HtrA is to deal with misfolded/damaged proteins in the periplasm. It can do this either by degrading (protease activity) or folding/capturing (chaperone/sequestering, C/S, activity) the aberrant protein. We investigated which of these functions are important to Typhimurium and . Point or deletion mutants of that encode variant HtrA molecules have been used in previous studies to investigate the role of different regions of HtrA in C/S and protease activity. These variants were placed under the control of the Typhimurium promoters and expressed in a Typhimurium mutant, GVB1343. Both wild-type HtrA and HtrA (HtrA S210A) lacking protease activity enabled GVB1343 to grow at high temperature (46 °C). Both molecules also significantly enhanced the growth/survival of GVB1343 in the liver and spleen of mice during infection. However, expression of wild-type HtrA enabled GVB1343 to grow to much higher levels than expression of HtrA S210A. Thus both the protease and C/S functions of HtrA operate during infection but the protease function is probably more important. Absence of either PDZ domain completely abolished the ability of HtrA to complement the growth defects of GVB1343 or .

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2009-03-01
2024-03-28
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