Abstract
The alterations in complex-type N-linked oligosaccharides that can occur when an animal cell line is transformed by two dissimilar viruses were examined by comparing the N-linked oligosaccharides of baby hamster kidney (BHK) cells, metabolically radiolabeled with [2-3H]mannose, to the same class of oligosaccharides from BHK cells separately transformed by Rous sarcoma virus (RS-BHK), an RNA retrovirus, and polyoma virus (PY-BHK), a DNA papovavirus. Based on experiments that utilized serial lectin affinity chromatography, glycosidase digestions, and methylation analyses, both RS-BHK and PY-BHK cells demonstrated a significant increase in the relative amounts of tri- and tetraantennary complex-type N-linked oligosaccharides containing the branching sequence, [GlcNAc-beta(1,6)Man-alpha(1,6)Man], compared to the nontransformed BHK cells. In addition, almost all of the poly-N-acetyllactosamine sequence, [GlcNAc-beta(1,3)-Gal-beta(1,4)], was expressed on the tri- and tetraantennary N-linked oligosaccharides from BHK and RS-BHK cells that contain the sequence, [GlcNAc-beta(1,6)Man-beta(1,6)Man]. The increase in the relative amounts of this latter sequence in the transformed cells, therefore, most likely results in an increase in the amount of poly-N-acetyllactosamine sequence on the N-linked glycopeptides of these cells. The analysis of the degree of sialylation of the complex-type N-linked oligosaccharides from BHK and RS-BHK cells by ion exchange chromatography revealed no apparent differences, and in both of these cell types approximately 3% of the glycopeptide fraction radiolabeled with mannose was recovered in a highly negatively charged fraction that was identified by keratanase digestion to be keratan sulfate.
Highlights
The criteria.Methylationanalysis of radiolabeled glycopeptide present studyutilized metabolically labeled glycopeptidesand fractions separated by serial lectin chromatography demon- lectin fractionation to confirm that compared to BHK cells, strated similar patterns of mannose substitution
QAE-chro- PY-BHK cells express quantitatively more of the antenna matography of three glycopeptide fractionsfrom both cell that bind to L-PHA.Since Rous sarcoma virus-transformed BHK cells (RS-BHK) cells show a similar types before and after mild acid treatment demonstrated a quantitative increase of these antenna, these results demonremarkably similarpattern of sialylation
Similar strate thattransformation of BHK cells by either an RNA or amounts of a fraction of the glycopeptides from both cells DNA tumor virus causes a similar phenotypic change in the that bound to no lectin was found to contain keratansulfate, ASN-linked oligosaccharides of these cells
Summary
Yamashita et al (6),who compared the Asn-linked oligosaccharides of BHK and PY-BHKcells by methylation analysis. BHK are similar in several respects, RS-BHK cells consist- Homogenates of PY-BHK cells have been shown to express ently demonstrated a %fold increase in the relative amounts about twice as much UDP-N-acetylg1ucosamine:a-mannoside of glycopeptides with the [Gal-P(1,4)GlcNAc-/3(1,6)Masne]- ~(1,6)-N-acetylglucosaminyltransferaascetivityas do BHK quence, those which bind to immobilized L-PHA. This in- cells, while the specific activities of the other five N-acetylcrease (Table I) has been expressed in terms of per cent of glucosaminyltransferaseswere similar (7). Van Den Eijnden, D., Joziasse, D., Koenderman, A., Blanken, W., Schiphorst, W., and Koppen, P. (1985) in Proceedings of theVIIIthInternationalSymposiumon Glycogonjugates, pp. 285-286, Praeger Press, New York
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