Expression of the core lipopeptide of the glycopeptidolipid surface antigens in rough mutants of Mycobacterium avium.
Toward studying the genetics, biosynthesis, and roles in the pathogenesis of the dominant surface glycopeptidolipid antigens of Mycobacterium avium, rough colony variants of M. avium serovar 2 were picked, cultured in quantity, and their lipid composition examined. Two of the rough (Rg) variants, Rg-3 and Rg-4, were devoid of glycopeptidolipids or any more elemental structures and thus were similar to those described previously. Two others, Rg-0 and Rg-1, each contained two novel lipopeptides, devoid of any of the carbohydrate substituents that confer serotypic activity on the glycopeptidolipids. The application of gas chromatography, fast atom bombardment-mass spectrometry and 1H NMR to lipopeptide I established the structure C32:2-beta-OH-fatty acyl-D-Phe-D-allo-Thr-D-Ala-L-alaninol. Lipopeptide II represented a minor variation of this structure: C32:1-beta-OH-fatty acyl-D-Phe-D-allo-Thr-D-Ala-L-alaninol. These newly discovered lipopeptides are identical to the fatty acyl-tripeptide-amino alcohol "core" component of the glycopeptidolipids of the M. avium complex, and thus the Rg-0 and Rg-1 variants represent a form of "deep rough" mutation in M. avium. Separately, we report that these rough variants of M. avium differ genetically from the smooth, virulent form by major deletions of portions of the genes responsible for glycopeptidolipid synthesis (Belisle, J. T., Klaczkiewicz, K., Brennan, P. J., Jacobs, W. R., Jr., and Inamine, J. M. (1993) J. Biol. Chem. 268, 10517-10523). The isolation of different sets of spontaneous mutants of M. avium differentially defective in the capacity to synthesize glycopeptidolipids provides the means to explore their biosynthesis and roles in opportunistic pathogenesis.
Belisle
J T
JT
Department of Microbiology, Colorado State University, Fort Collins 80523.
McNeil
M R
MR
Chatterjee
D
D
Inamine
J M
JM
Brennan
P J
PJ
D000595:Amino Acid Sequence; D000942:Antigens, Bacterial / Q000096:biosynthesis / Q000737:chemistry / Q000302:isolation & purification; D000954:Antigens, Surface / Q000096:biosynthesis / Q000737:chemistry / Q000302:isolation & purification*; D002855:Chromatography, Thin Layer; D008401:Gas Chromatography-Mass Spectrometry; D014644:Genetic Variation; D006001:Glycoconjugates / Q000096:biosynthesis / Q000737:chemistry / Q000302:isolation & purification*; D009682:Magnetic Resonance Spectroscopy; D008969:Molecular Sequence Data; D009162:Mycobacterium avium / Q000235:genetics / Q000254:growth & development / Q000378:metabolism*; D016339:Spectrometry, Mass, Fast Atom Bombardment
D000942:Antigens, Bacterial; D000954:Antigens, Surface; D006001:Glycoconjugates