Sodium butyrate causes reexpression of three membrane proteins on glycolipid-anchoring mutants.
Murine Thy-1-negative lymphoma mutants synthesize membrane proteins that normally bear glycolipid anchors but do not express these proteins on the cell surface. This phenotype may reflect altered regulation of gene(s) required for anchor biosynthesis. Since tissue culture cells treated with sodium butyrate transcribe new DNA sequences and since these transcripts are translated, it was of interest to determine whether butyrate treatment could restore surface expression of lipid-anchored proteins. When Thy-1-negative lymphoma mutants (complementation groups A-C, E, F, and H) were cultured for three days in 1.5 mM butyrate, a small percentage of the class H cells acquired phosphatidylinositol-specific phospholipase C-releasable surface Thy-1 and J11d. Membrane-associated Thy-1 was not observed before 24 h of treatment. Induction was reversible. Cell fusion studies have shown that murine LM (TK-) fibroblasts can be assigned to the class H lymphoma complementation group. Although these cells synthesize Ly-6, this normally lipid-anchored protein is absent from the cell surface. When LM (TK-) cells were cultured for three days in butyrate, 10% of the cells reversibly expressed Ly-6. In addition, LM (TK-) cells transfected with a plasmid encoding Thy-1 do not express Thy-1, but could be induced to express both Ly-6 and Thy-1 by butyrate treatment. Northern analysis of total RNA from Ly-6/Thy-1-expressing cells indicates that increased steady-state transcript levels cannot account for surface expression of these proteins. We conclude that the lack of expression of three proteins at the surface of class H mutant and the LM (TK-) cells is not due to gross structural lesions in genes along the anchor biosynthetic pathway.
Scripps Clinic and Research Foundation, La Jolla, California 92037.
E J
EJ
Tisdale
J C
JC
Schimenti
A M
AM
Tartakoff
D000818:Animals; D000950:Antigens, Ly / Q000235:genetics; D000954:Antigens, Surface / Q000235:genetics; D002087:Butyrates / Q000494:pharmacology*; D020148:Butyric Acid; D002460:Cell Line; D005347:Fibroblasts / Q000187:drug effects / Q000378:metabolism; D015870:Gene Expression / Q000187:drug effects*; D006017:Glycolipids / Q000235:genetics* / Q000276:immunology; D008223:Lymphoma / Q000235:genetics / Q000473:pathology; D008562:Membrane Glycoproteins / Q000235:genetics* / Q000276:immunology; D051379:Mice; D009154:Mutation; D018800:Thy-1 Antigens; D013937:Thymidine Kinase; D013945:Thymoma / Q000235:genetics / Q000473:pathology; D014407:Tumor Cells, Cultured
D000950:Antigens, Ly; D000954:Antigens, Surface; D002087:Butyrates; D006017:Glycolipids; D008562:Membrane Glycoproteins; D018800:Thy-1 Antigens; D020148:Butyric Acid; D013937:Thymidine Kinase