Identification of a PEST‐like motif in listeriolysin O required for phagosomal escape and for virulence in Listeria monocytogenes

Molecular Microbiology - Tập 39 Số 5 - Trang 1124-1139 - 2001
M A Lety1, Claude Fréhel1, Iharilalao Dubail1, Jean‐Luc Béretti1, Samer Kayal1, Patrick Berche1, Alain Charbit2
1INSERM U‐411, CHU Necker‐Enfants Malades, 156 rue de Vaugirard, 75730 Paris Cedex 15, France.
2*For correspondence. E-mail charbit@ necker.fr; Tel. (+33) 1 40 61 53 76; Fax (+33) 1 40 61 55 92.

Tóm tắt

The hly‐encoded listeriolysin O (LLO) is a major virulence factor secreted by the intracellular pathogen Listeria monocytogenes, which plays a crucial role in the escape of bacteria from the phagosomal compartment. Here, we identify a putative PEST sequence close to the N‐terminus of LLO and focus on the role of this motif in the biological activities of LLO. Two LLO variants were constructed: a deletion mutant protein, lacking the 19 residues comprising this sequence (residues 32–50), and a recombinant protein of wild‐type size, in which all the P, E, S or T residues within this motif have been substituted. The two mutant proteins were fully haemolytic and were secreted in culture supernatants of L. monocytogenes in quantities comparable with that of the wild‐type protein. Strikingly, both mutants failed to restore virulence to a hly‐negative strain in vivo. In vitro assays showed that L. monocytogenes expressing the LLO deletion mutant was strongly impaired in its ability to escape from the phagosomal vacuole and, subsequently, to divide in the cytosol of infected cells. This work reveals for the first time that the N‐terminal portion of LLO plays an important role in the development of the infectious process of L. monocytogenes.

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Molecular Microbiology

Tập 39 Số 5

1124-1139

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