Isolation and preliminary characterisation of DNA-protein complexes from the mitochondria of Saccharomyces cerevisiae

Four independent rat L6 myoblast cell lines have been selected in a single step for resistance to the cytotoxic effects of the lectin concanavalin A (conA). In contrast to parental wild-type myoblast lines, all of the variant clones are unable to undergo normal cellular differentiation to form multinucleated myotubes or biochemical differentiation to produce an increase in the specific activity of the muscle-specific enzyme, creatine phosphokinase (CPK). The correlation between lectin resistance and loss of fusion potential is very tight; clonal variation studies show that there is less than a 2.8×10^?8 chance that the two are not directly related. Membrane preparations from the conA-resistant myoblast lines incorporate significantly less GDP-¹⁴C]mannose into the lipid intermediates of protein glycosylation than preparations from parental wild-type cells. Also, conversion of mannose label to fucose occurs in myoblasts and this pathway is more active in conA-resistant cells than wild-type cells. Reduced binding of labelled conA to the cell surfaces of variant myoblasts was observed which may result from alterations to membrane glycoprotein receptors. These studies suggest that mannosylated glycoproteins of the cell surface play a role in the development of the myotubes from myoblasts. Lectin-resistant myoblasts should be useful model systems for investigating what appears to be a pleiotropic mutation affecting the myogenesis process through membrane modifications.