Effect of bacitracin on flagellar assembly and presumed glycosylation of the flagellins of Methanococcus deltae

• Published in: Archives of microbiology 1993-09, Vol.160 (3), p.179-185
• Main Authors: BAYLEY, D. P, KALMOKOFF, M. L, JARRELL, K. F
• Format: Article
• Language: English
• Subjects: Bacteriology; Biogenesis of cell structures, supramolecular organization; Biological and medical sciences; Biotechnology; Fundamental and applied biological sciences. Psychology; Microbiology
• ISSN: 0302-8933; 1432-072X
• DOI: 10.1007/BF00249122

Methanococcus deltae is an irregularly-shaped coccoid methanogen which possesses peritrichously arranged flagella. The flagella are composed of 2 flagellins of M sub(r) = 27000 and 32000 and possess a carbohydrate component as determined by thymol-sulfuric acid staining of SDS-PAGE. Cell growth was sensitive to bacitracin at levels near 10 mu g/ml. Growth in the presence of 5 mu g/ml bacitracin resulted in the appearance of presumably hypoglycosylated flagellins as detected by Western immunoblotting. Continual passage of cells from 5 mu g/ml bacitracin through 10, 25, and 50 mu g/ml bacitracin resulted in cells capable of growth at 100 mu g/ml bacitracin. Western immunoblot analysis revealed that cells grown in the highest concentration of bacitracin no longer possessed native flagellins but only the hypoglycosylated forms. Electron microscopy corroborated the absence of normal flagella. These studies suggest that a bacitracin-sensitive dolichol-diphosphate carrier is responsible for attachment of at least a large proportion of the carbohydrate content of the flagellins, and that a minimum amount of glycosylation is essential for normal flagellum assembly.