Biochemical characterization of mIgM- variants of the murine B-cell lymphoma, WEHI 279.1.
The membrane immunoglobulin M (mIgM) of a B lymphocyte serves as a receptor for its cognate antigen. Our aim is to elucidate the structure and function of this membrane-bound receptor. The first step is to determine the requirements for proper membrane placement of IgM. We have used mIgM-positive B lymphocyte tumors from which we isolated mIgM negative variants by immunoselection. We report here the initial characterization of mIgM- variants isolated by repeated cycles of selection of the murine B lymphoma, WEHI 279.1, with goat anti-mouse immunoglobulin (G alpha MIg) and complement. These particular variants were chosen from a pool of more than 150 variants originally isolated because they resulted from several selection schemes and clearly had different origins. By analysis of their proteins, we have found three major phenotypes that do not produce mIgM: reduced microns, microseconds and L chain levels within cells, loss of microns and microseconds but retention of L chain synthesis, and loss of microns but retention of reduced amounts of microseconds and L chain. The defects underlying these phenotypes produce complex changes in the synthesis, turnover, and secretion of the mu or L chains involved. We performed experiments comparing the effects of the glycosylation inhibitor tunicamycin on variants with reduced mu and L levels with its effects on variants with L but no mu chains. These experiments suggested that mu and L chain synthesis are controlled coordinately at the level of protein synthesis. We have not yet isolated any variants lacking L chain synthesis or any appearing to have gross structural defects in the micron protein. This analysis is the first phase of the detailed characterization of the requirements for proper synthesis, processing, tetramer formation, and membrane display of mIgM on B lymphoma tumors in mice.
D000818:Animals; D001402:B-Lymphocytes / Q000276:immunology; D002460:Cell Line; D002999:Clone Cells; D014644:Genetic Variation; D007075:Immunoglobulin M / Q000235:genetics*; D008223:Lymphoma / Q000235:genetics / Q000276:immunology*; D051379:Mice; D010641:Phenotype
D007075:Immunoglobulin M