Protein glycosylation in heat-sensitive and thermotolerance-deficient mutants of Chinese hamster ovary cells.
Chinese hamster ovary (CHO) cells are capable of developing a high degree of thermotolerance in response to appropriate heat conditioning. In this study we examined the relationship between thermotolerance development and protein glycosylation using four sublines of CHO cells. Two of these CHO sublines are characterized by an increased heat sensitivity and impaired cellular capacity for thermotolerance development. The data show that thermotolerance development after heat conditioning in the heat-sensitive, thermotolerance-deficient mutants was accompanied by reduced labeling of a Mr 50,000 glycoprotein (GP50), in both soluble and insoluble cell fractions. Similarly, activation of UDP-N-acetylgalactosaminyltransferase (Gal-NAcT) after hyperthermia was almost completely abolished in these cell lines. Both of these endpoints have been correlated previously with thermotolerance expression. The data are consistent with the glycosylation hypothesis that attributes increased heat resistance of thermotolerant cells, at least in part, to enhanced glycosylation and accumulation of endogenous glycoproteins, such as GP50.
Henle
K J
KJ
Department of Medicine, University of Arkansas for Medical Sciences, Little Rock 72205.
Nagle
W A
WA
Bedford
J S
JS
Harvey
W F
WF
D000818:Animals; D002460:Cell Line; D002470:Cell Survival; D004591:Electrophoresis, Polyacrylamide Gel; D005700:Galactosyltransferases / Q000378:metabolism; D006023:Glycoproteins / Q000096:biosynthesis / Q000378:metabolism*; D006031:Glycosylation; D006358:Hot Temperature; D009154:Mutation; D017350:N-Acetylgalactosaminyltransferases; D010779:Photofluorography
D006023:Glycoproteins; D005700:Galactosyltransferases; D017350:N-Acetylgalactosaminyltransferases; C003742:UDPgalactosamine-galactose acetylgalactosaminyltransferase