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Molecular Genetics and Immunobiology of Mycobacteria |
Institut de Pharmacologie et de Biologie Structurale, Unité Mixte de Recherche du Centre de National de Recherche Scientifique et de l’Université Paul Sabatier (UMR 5089), 205 route de Narbonne, 31077 Toulouse cedex 04, France1
Department of Microbiology and Immunology, University of Melbourne, Victoria 3010, Australia2
Institut d’Exploration Fonctionnelle des Génomes (IFR 109), 118 route de Narbonne, 31062 Toulouse cedex, France3
Author for correspondence: Mamadou Daffé. Tel: +33 561 175 569. Fax: +33 561 175 580. e-mail: mamadou.daffe{at}ipbs.fr
Glycopeptidolipids (GPLs) are a class of species- or type-specific mycobacterial lipids and major constituents of the cell envelopes of many non-tuberculous mycobacteria. To determine the function of GPLs in the physiology of these bacteria, a mutant of Mycobacterium smegmatis in which the gene encoding a mycobacterial nonribosomal peptide synthetase has been inactivated by transposon mutagenesis was analysed. Labelling experiments indicated that half of the bacterial GPLs were located on the cell surface and represented 85% of the surface-exposed lipids of the parent strain whereas the mutant was defective in the production of the GPLs. Compared to the parent smooth morphotype strain, the GPL-deficient mutant strain exhibited a rough colony morphology, an increase of the cell hydrophobicity and formed huge aggregates. As a consequence, the mutant cells were no longer able to bind ruthenium red, as observed by transmission electron microscopy. The altered surface properties of the mutant cells also affected the phagocytosis of individual bacilli by human monocyte-derived macrophages since mutant cells were internalized more rapidly than cells from the parent strain. Nevertheless, no specific release of surface constituents into the culture broth of the mutant was observed, indicating that the cell surface is composed of substances other than GPLs and that these are essential for maintaining the architecture of the outermost layer of the cell envelope. Importantly, the absence of these major extractable lipids of M. smegmatis from the mutant strain has a profound effect on the uptake of the hydrophobic chenodeoxycholate by cells, indicating that GPLs are involved in the cell wall permeability barrier of M. smegmatis. Altogether, these data showed that, in addition to being distinctive markers of numerous mycobacterial species, GPLs play a role in the bacterial phenotype, surface properties and cell wall permeability.
Keywords: mycobacteria, cell wall permeability, morphology, ultrastructure, glycolipids
Abbreviations: GPL, glycopeptolipid; MDM, monocyte-derived macrophage; OL, outer layer; SEM, surface-exposed material
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