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JBC, Vol. 252, Issue 3, 1107-1116, Feb, 1977
E. B. Briles, E. Li and S. Kornfeld
Several clones of Chinese hamster ovary cells have been selected for their
resistance to the toxic effects of wheat germ agglutinin. The clones do not
bind wheat germ agglutinin as well as parent cells and are 5- to 250-fold
more resistant to the toxic effects of the lectin. Of three clones studied
in detail, all exhibit a decrease in wheat germ agglutinin binding
affinity. Two have normal numbers of wheat germ agglutinin binding sites,
while one (Clone 13) has a 65% decrease in binding sites. Crude membrane
preparations of the clones have a decrease in sialic acid content relative
to parent cells, and Clone 13 membranes are also deficient in galactose,
while the mannose and hexosamine contents of all three clones are normal.
The membrane sugar deficiencies affect both glycoproteins and glycolipids.
Sialyl-lactosylceramide is the major glycolipid in parent cells, while
Clones 1 and 1021 have lactosylceramide and Clone 13 has glucosylceramide
as the predominant glycolipid. Labeling experiments with
N-[G-3H]acetylmannosamine suggest that Clone 1021 cells have a block in the
transfer of sialic acid from CMP-sialic acid to glycoprotein and glycolipid
acceptors. Yet CMP-sialic acid:glycoprotein sialyl-transferase activity in
cell lysates of Clone 1021 cells is 80% of normal. While CMP-sialic
acid:lactosylceramide sialyl-transferase activity is only 25% of normal, it
can be restored to normal or elevated levels by sodium butyrate induction
without an associated increase in cellular sialyl-lactosylceramide content.
Similarly, the galactose-deficient Clone 13 can synthesize UDP-galactose
and has normal levels of UDP-galactose:glycoprotein galactosyltransferase
and UDP-galactose:glucosylceramide galactosyltransferase when assayed in
vitro. The glycosyltransferases of both these clones can utilize their own
glycoproteins as sugar acceptors in in vitro assays. These data suggest
that the variant cells fail to carry out specific glycosyltransferase
reactions in vivo despite the fact that they possess the appropriate
nucleotide sugars, glycoprotein and glycolipid acceptors, and
glycosyltransferases.
Isolation of wheat germ agglutinin-resistant clones of Chinese hamster ovary cells deficient in membrane sialic acid and galactose
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