UDP-N-acetylglucosamine:dolichyl-phosphate N-acetylglucosamine-1-phosphate transferase is amplified in tunicamycin-resistant soybean cells.
A tunicamycin-resistant soybean cell line was developed by gradually increasing the concentration of tunicamycin in the growth medium. At the final stage, the resistant cells could survive in media containing 60 micrograms/ml of tunicamycin, whereas normal cells show a greatly retarded growth rate at 0.5 microgram/ml of antibiotic. The tunicamycin-resistant cells had a greater than 40-fold increase in the activity of the enzyme UDP-GlcNAc:dolichyl-P GlcNAc1P transferase, a 2-3-fold increase in the activity of dolichyl-P-mannose synthase, but no increase in the activities of other enzymes of the lipid-linked saccharide pathway such as dolichyl-P-glucose synthase or mannosyl transferases. There was also no change in the activities of the glycoprotein-processing enzymes, glucosidase I or glucosidase II, as compared to wild-type cells. The increase in GlcNAc1P transferase was due to an increased production of enzyme, as seen by a dramatic increase in the amount of a 39-kDa protein, which is presumed to be this enzyme protein. The GlcNAc1P transferase from tunicamycin-resistant cells was equally sensitive to tunicamycin as was the wild-type enzyme, but was considerably more labile to temperatures above 30 degrees C. The activity in tunicamycin-resistant cells was greatly stimulated by exogenous dolichyl-P. The spectrum of oligosaccharides from labeled lipid-linked oligosaccharides was similar in wild-type and tunicamycin-resistant soybean cells, but the resistant cells had significantly greater amounts of the shorter and much lower amounts of the larger-sized oligosaccharides.
Zeng
Y
Y
Department of Biochemistry and Molecular Biology, University of Arkansas for Medical Sciences, Little Rock 72205, USA.
Elbein
A D
AD
D000900:Anti-Bacterial Agents / Q000494:pharmacology*; D002460:Cell Line; D004352:Drug Resistance, Microbial; D006031:Glycosylation; D013025:Glycine max; D017855:Transferases (Other Substituted Phosphate Groups) / Q000378:metabolism*; D014415:Tunicamycin / Q000494:pharmacology*
D000900:Anti-Bacterial Agents; D014415:Tunicamycin; D017855:Transferases (Other Substituted Phosphate Groups); C023222:UDPacetylglucosamine-dolichyl-phosphate acetylglucosamine-1-phosphate transferase