Exploring the role of the CTL epitope region of listeriolysin O in the pathogenesis of Listeria monocytogenes

Microbiology
Marie-Annick LetyAlain Charbit

Abstract

Listeria monocytogenes is a facultative intracellular bacterial pathogen responsible for severe opportunistic infections in humans and animals. The secreted cholesterol-dependent cytolysin, listeriolysin O (LLO), mediates phagosomal escape and allows bacterial growth in the cytosol of infected cells. In order to identify new LLO determinants participating in bacterial pathogenesis, this study focused on a major target of LLO proteolytic cleavage in vitro, the CTL epitope region (residues 91-99). Mutations were generated by site-directed mutagenesis in the epitope or in the two clusters of positive charges flanking the epitope. Two LLO mutants (a single mutation K103A and a double mutation R89G, K90G) were normally and stably secreted by L. monocytogenes. In contrast, a mutant carrying four amino acid substitutions in the epitope itself (Y92K, D94A, E97K, Y98F) was highly susceptible to proteolytic degradation. While these three LLO mutant proteins showed a reduced haemolytic activity, they all promoted efficient phagosomal escape and intracellular multiplication in different cell types, and were non-cytotoxic. The deletion of the epitope (Delta91-99), as well as the substitution of two, three or four of the four lysine residues...Continue Reading

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Citations

May 24, 2008·Infection, Genetics and Evolution : Journal of Molecular Epidemiology and Evolutionary Genetics in Infectious Diseases·Renato H OrsiMartin Wiedmann
Aug 31, 2007·International Journal of Food Microbiology·Dongyou LiuFrank W Austin
Feb 13, 2013·Human Vaccines & Immunotherapeutics·Rui Sun, Yuqin Liu
Jan 9, 2014·PloS One·Leigh A KnodlerOlivia Steele-Mortimer
Jun 2, 2012·Trends in Microbiology·Mélanie Anne HamonPascale Cossart

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