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Applied and Environmental Microbiology, December 2005, p. 8314-8322, Vol. 71, No. 12
0099-2240/05/$08.00+0     doi:10.1128/AEM.71.12.8314-8322.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

Listeria monocytogenes PerR Mutants Display a Small-Colony Phenotype, Increased Sensitivity to Hydrogen Peroxide, and Significantly Reduced Murine Virulence

Department of Microbiology, University College Cork, Cork, Ireland,¹ Alimentary Pharmabiotic Centre, University College Cork, Cork, Ireland²

Received 26 April 2005/ Accepted 11 August 2005

Deletion of perR in Listeria monocytogenes results in a small-colony phenotype (perR_sm) that is slow growing and exhibits increased sensitivity to H₂O₂. At a relatively high frequency, large-colony variants (perR_lg) arise, which are more resistant to H₂O₂ than the wild-type and ultimately dominate the culture. Transcriptional analysis revealed that the kat gene (catalase) is up-regulated in both types of mutants and that the highest level is apparent in perR_sm mutants, demonstrating PerR regulation of this gene. Overexpression of the catalase gene in the wild-type background resulted in a slower-growing strain with a smaller colony size similar to that of perR_sm. By combining a bioinformatic approach with experimental evidence, other PerR-regulated genes were identified, including fur, lmo0641, fri, lmo1604, hemA, and trxB. The transcriptional profile of these genes in both mutant backgrounds was similar to that of catalase in that a higher level of expression was observed in perR_sm than in the wild type or perR_lg. Murine studies revealed that the virulence potential of the perR_sm mutant is substantially reduced compared to that of the wild-type and perR_lg strains. Collectively, the data demonstrate that the perR_sm mutant represents the true phenotype associated with the absence of PerR, which is linked to overexpression of regulated genes that negatively affect bacterial homeostasis both in vitro and in vivo. A subsequent secondary mutation occurred at a high frequency, which resulted in phenotypic reversion to a large-colony phenotype with increased fitness that may have obstructed the analysis of the role of PerR in the physiology of the bacterial cell.

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