Abstract
Methanococcus deltae is an irregularly-shaped coccoid methanogen which possesses peritrichously arranged flagella. The flagella are composed of 2 flagellins of M 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 μg/ml. Growth in the presence of 5μg/ml bacitracin resulted in the appearance of presumably hypoglycosylated flagellins as detected by Western immunoblotting. Continual passage of cells from 5 μg/ml bacitracin through 10, 25, and 50 μg/ml bacitracin resulted in cells capable of growth at 100 μg/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 carbonhydrate content of the flagellins, and that a minimum amount of glycosylation is essential for normal flagellum assembly.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Balch WE, Fox GE, Magrum LJ, Woese CR, Wolfe RS (1979) Methanogens. reevaluation of a unique biological group. Microbiol Rev 43: 260–296
Faguy DM, Koval SF, Jarrell KF (1992) Correlation between glycosylation of flagellin proteins and sensitivity of flagellar filaments to Triton X-100 in methanogens. FEMS Microbiol Lett 90: 129–134
Gerl L, Sumper M (1988) Halobacterial flagellins are encoded by a multigene family. Characterization of the five flagellin genes. J Biol Chem 263: 13246–13251
Harris JE, Pinn PA (1985) Bacitracin-resistant mutants of a mesophilic Methanobacterium species. Arch Microbiol 143: 151–153
Hilpert R, Winter J, Hammes W, Kandler O (1981) The sensitivity of archaebacteria to antibiotics. Zentbl Bakteriol Hyg I Abt Orig C2: 11–20
Jain MK, Zeikus G (1987) Methods for isolation of auxotrophic mutants of Methanobacterium ivanovii and initial charcterization of acetate auxotrophis. Appl Environ Microbiol 53: 1387–1390
Jansen JJM, Mulder WR, DeCaluwé GLJ, Vlak JM, DeGrip WJ (1991) In vitro expression of bovine opsin using recombinant baculovirus: the role ofglutamic acid (134) in opsin biosynthesis and glycosylation. Biochim Biophys Acta 1089: 68–76
Kalmokoff ML, Jarrell KF (1991) Cloning and sequencing of a multigene family encoding the flagellins of Methanococcus voltae. J Bacteriol 173: 7113–7125
Kalmokoff ML, Jarrell KF, Koval SF (1988) Isolation of flagella from the archebacterium Methanococcus voltate by phase separation with Triton X-114. J Bacteriol 170: 1752–1758
Kalmokoff ML, Karnauchow TM, Jarrell KF (1990) Conserved N-terminal sequences in the flagellins of archaebacteria. Biochem Biophys Res Commun 167: 154–160
Kalmokoff ML, Koval SF, Jarrell KF (1992) Relatedness of the flagellins from methanogens. Arch Microbiol 157: 481–487
Koval SF, Jarrell KF (1987) Ultrastructure and biochemistry of the cell wall of Methanococcus voltae. J Bacteriol 169: 1298–1306
Küpcü Z März L, Messner P, Sleytr UB (1984) Evidence for the glycoprotein nature of the crystalline cell wall surface layer of Bacillus stearothermophilus strain NRS2004/3a. FEBS Lett 173: 185–190
Ladapo J, Whitman WB (1990) Method for isolation of auxotrophs in the methanogenic archaebacteria: role of the acetyl-CoA pathway of autotrophic CO2 fixation in Methanococcus maripaludis. Proc Natl Acad Sci USA 87: 5598–5602
Laemmli UK (1970) Cleavage of structure proteins during the assembly of the head of bacteriophage T4. Nature 227: 680–685
Lechner J, Wiedland F (1980) Structure and biosythesis of prokaryotic glycoproteins. Annu Rev Biochem 58: 173–194
Lechner J, Wieland F, Sumper M (1985) Transient methylation of dolichyl oligosaccharides is an obligatory step in halobacterial sulfated glycoprotein biosynthesis. J Biol Chem 260: 8984–8989
Mescher MF, Strominger JL (1978) Glycosylation of the surface glycoprotein of Halobacterium salinarium via a cyclic pathway of lipid-linked intermediates. FEBS Lett 89: 37–41
Messner P., Sleytr UB (1991) Bacterial surface layer glycoproteins. Glycobiology 1: 545–551
Pang S-Z, Rasmussen J, Ye G-N, Sanford JC (1992) Use of the signal peptide of Pisum vicilin to translocate β-glucuronidase in Nicotiana tabacum. Gene 112: 229–234
Racusen D (1979) Glycoprotein detection in polyacrylamide gel with thymol and sulfuric acid. Anl Biochem 99: 474–476
Reisinger P, Seidel H, Tschesche H, Hammes WP (1980) The effect of nisin on murein synthesis. Arch Microbiol 127: 187–193
Romesser JA, Wolf RS Mayer F, Spiess E, Walther-Mauruschat A (1979) Methanogenium, a new genus of marine methanogenic bacteria, and characterization of Methanogenium cariaci sp. nov. and Methanogenium marisnigri sp. nov. Arch Microbiol 121: 147–153
Siewert G, Strominger JL (1967) Bacitracin: an inhibitor of the dephosphorylation of lipid pyrophosphate, an intermediate in biosynthesis of the peptidoglycan of bacterial cell walls. Proc Natl Acad Sci USA 57: 767–773
Sprott GD, Koval SF, Schnaitman CA (1993) Cell fractionation. In: Gerhardt P (ed) Methods for general and molecular bacteriology American Society for Microbiology, Washington DC (in press)
Sumper M (1987) Halobacterial glycoprotein biosynthesis. Biochim Biophys Acta 906: 69–79
Sumper M, Herrman G (1978) Studies on the biosynthesis of bacterio-opin: demonstration of the existence of protein species structurally related to bacterio-opsin. Eur J Biochem 89: 229–235
Towbin M, Staehelin T, Gordon J (1979) Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci USA 76: 4350–4354
Wieland F, Lechner J, Berhardt G, Sumper M (1981) Sulphation of a repetitive saccharide in halobacterial cell wal glycoprotein. FEBS Lett 132: 319–323
Wieland F, Lechner J, Sumper M (1982) The cell wall glycoprotein of halobacteria: structural, functional and biosynthetic aspects. Zentbl Bakterio Hyg I Abt Orig C 3: 161–170
Whitman WB, (1989) Methanococcales. In: Staley JT, Bryant MP, Pfennig N (eds) Bergey's manual of systematic bacteriology, vol 3. Williams and Wilkins, Baltimore, Md., pp 2185–2190
Whitman WB, Bowen TL, Boone DR (1992) The methanogenic bacteria. In: Balows A, Trüper HG, Dworkin M, Harder W, Schleifer K-H (eds) The prokaryotes, vol 1 Springer, New York Berlin Heidelberg, pp 719–767
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Bayley, D.P., Kalmokoff, M.L. & Jarrell, K.F. Effect of bacitracin on flagellar assembly and presumed glycosylation of the flagellins of Methanococcus deltae . Arch. Microbiol. 160, 179–185 (1993). https://doi.org/10.1007/BF00249122
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00249122