Changes of nuclear glycoproteins in normal and simian virus 40-transformed WI-38 human lung fibroblasts

To investigate changes in nuclear glycoproteins after transformation, nuclei and nuclear extracts were prepared from the normal and SV40-transformed WI-38 human lung fibroblasts grown in vitro. The nuclei of the normal and transformed cells were isolated by a vigorous pipetting method, and the isolated nuclei were verified by morphological and biochemical examinations. The nuclear glycocomponents, which were labelled with fluorescein isothiocyanate-lectins, were evenly found all through the isolated nuclei. Lectin blotting analysis showed that dozens of nuclear proteins contain both Man and GlcNAc moieties. The glycoproteins bearing the terminal N-acetylglucosamine sugar moieties were also demonstrated by in vitro galactosylation with [3H]-galactose. The result showed that nine glycoproteins specifically appeared or disappeared upon viral transformation or as SV40 replication products, indicating that there are significant differences in nuclear glycoproteins between the normal and SV40-transformed cells. These results suggest that the modification of Man and GlcNAc moieties are dynamic changes.