Monoclonal antibody to the amino-terminal L sequence of murine leukemia virus glycosylated gag polyproteins demonstrates their unusual orientation in the cell membrane.
To analyze cell surface murine leukemia virus gag protein expression, we have prepared monoclonal antibodies against the spontaneous AKR T lymphoma KKT-2. One of these antibodies, 43-13, detects an AKR-specific viral p12 determinant. A second monoclonal antibody, 43-17, detects a novel murine leukemia virus-related antigen found on glycosylated gag polyproteins (gp95gag, gp85gag, and gp55gag) on the surface of cells infected with and producing ecotropic endogenous viruses, but does not detect antigens within these virions. The 43-17 antibody immunoprecipitates the precursor of the cell surface gag protein whether in its glycosylated or unglycosylated state, but does not detect the cytoplasmic precursor of the virion gag proteins (Pr65gag). Based on these findings, we have localized the 43-17 determinant to the unique amino-terminal part of the glycosylated gag polyprotein (the L domain). We have determined that gp95gag contains L-p15-p12-p30-p10 determinants, whereas gp85gag lacks the carboxyterminal p10 determinant, and gp55gag lacks both p30 and p10 carboxy terminal determinants. Analysis of cell surface gag expression with the 43-17 antibody leads us to propose that the L domain plays a crucial role in (i) the insertion and orientation of murine leukemia virus gag polyproteins in the cell membrane and (ii) the relative abundance of expression of AKR leukemia virus versus Moloney murine leukemia virus glycosylated gag polyproteins in infected cells.
E A
EA
Pillemer
D A
DA
Kooistra
O N
ON
Witte
I L
IL
Weissman
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D000914:Antibodies, Viral / Q000276:immunology; D000918:Antibody Specificity; D002462:Cell Membrane / Q000276:immunology / Q000648:ultrastructure; D002478:Cells, Cultured; D011232:Chemical Precipitation; D000939:Epitopes; D015683:Gene Products, gag; D006023:Glycoproteins / Q000276:immunology; D009052:Leukemia Virus, Murine / Q000276:immunology*; D008223:Lymphoma / Q000276:immunology; D008565:Membrane Proteins / Q000276:immunology*; D051379:Mice; D011498:Protein Precursors / Q000276:immunology; D012191:Retroviridae Proteins / Q000276:immunology*; D014771:Virion / Q000276:immunology
D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral; D000939:Epitopes; D015683:Gene Products, gag; D006023:Glycoproteins; D008565:Membrane Proteins; D011498:Protein Precursors; D012191:Retroviridae Proteins