1887

Abstract

Mycoloyltransferases (Myts) play an essential role in the biogenesis of the cell envelope of members of the , a group of bacteria that includes the mycobacteria and corynebacteria. While the existence of several functional genes has been demonstrated in both mycobacteria and corynebacteria (), the disruption of any of these genes has at best generated cell-wall-defective but always viable strains. To investigate the importance of Myts on the physiology of members of the , a double mutant of was constructed by deleting and , and the consequences of the deletion on the viability of the mutant, the transfer of corynomycoloyl residues onto its cell-wall arabinogalactan and trehalose derivatives, and on its cell envelope ultrastructure were determined. The double mutant strain failed to grow at 34 °C and exhibited a growth defect and formed segmentation-defective cells at 30 °C. Biochemical analyses showed that the double mutant elaborated 60 % less cell-wall-bound corynomycolates and produced less crystalline surface layer proteins associated with the cell surface than the parent and -inactivated mutant strains. Freeze-fracture electron microscopy showed that the Δ Δ double mutant, unlike the wild-type and -inactivated single mutant strains, frequently exhibited an additional fracture plane that propagated within the plasma membrane and rarely exposed the S-layer protein. Ultra-thin sectioning of the double mutant cells showed that they were totally devoid of the outermost layer. Complementation of the double mutant with the wild-type or gene restored completely or partially this phenotype. The data indicate that Myts are important for the physiology of and reinforce the concept that these enzymes would represent good targets for the discovery of new drugs against the pathogenic members of the .

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2004-01-01
2024-12-07
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