Inactivation of the antigen 85C gene profoundly affects the mycolate content and alters the permeability of the Mycobacterium tuberculosis cell envelope.
The antigen 85 complex of Mycobacterium tuberculosis consists of three abundantly secreted proteins. The recent characterization of a mycoloyltransferase activity associated in vitro with each of these antigens suggested that they are potentially important for the building of the unusual cell envelope of mycobacteria. To define the physiological role of these proteins, the gene coding for antigen 85C was inactivated by transposon mutagenesis. The resulting mutant was shown to transfer 40% fewer mycolates to the cell wall with no change in the types of mycolates esterifying arabinogalactan or in the composition of non-covalently linked mycolates. As a consequence, the diffusion of the hydrophobic chenodeoxycholate and the hydrophilic glycerol, but not that of isoniazid, was found to be much faster through the cell envelope of the mutant than that of the parent strain. Taken together, these data demonstrate that: (i) antigen 85C is involved directly or indirectly in the transfer of mycolates onto the cell wall of the whole bacterium; (ii) the enzyme is not specific for a given type of mycolate; and (iii) the cell wall-linked mycolate layer may represent a barrier for the diffusion of small hydrophobic and hydrophilic molecules.
Jackson
M
M
Unité de Génétique Mycobactérienne, Institut Pasteur, Paris, France.
Raynaud
C
C
Lanéelle
M A
MA
Guilhot
C
C
Laurent-Winter
C
C
Ensergueix
D
D
Gicquel
B
B
Daffé
M
M
D000217:Acyltransferases; D000818:Animals; D000942:Antigens, Bacterial / Q000235:genetics*; D015139:Blotting, Southern; D015153:Blotting, Western; D001853:Bone Marrow / Q000378:metabolism; D002463:Cell Membrane Permeability; D002635:Chenodeoxycholic Acid / Q000378:metabolism; D002855:Chromatography, Thin Layer; D004251:DNA Transposable Elements; D015180:Electrophoresis, Gel, Two-Dimensional; D004591:Electrophoresis, Polyacrylamide Gel; D005786:Gene Expression Regulation; D015723:Gene Library; D005990:Glycerol / Q000378:metabolism; D008081:Liposomes / Q000378:metabolism; D008264:Macrophages / Q000378:metabolism; D008563:Membrane Lipids / Q000378:metabolism; D051379:Mice; D008807:Mice, Inbred BALB C; D008826:Microbial Sensitivity Tests; D008957:Models, Genetic; D016296:Mutagenesis; D009169:Mycobacterium tuberculosis / Q000235:genetics*; D009171:Mycolic Acids / Q000378:metabolism*; D016622:Silver Staining; D013997:Time Factors
D000942:Antigens, Bacterial; D004251:DNA Transposable Elements; D008081:Liposomes; D008563:Membrane Lipids; D009171:Mycolic Acids; D002635:Chenodeoxycholic Acid; D000217:Acyltransferases; C119774:antigen 85C, Mycobacterium tuberculosis; D005990:Glycerol