FIG. 2.

Expression of human cyto-LysRS and mito-LysRS in yeast. Total extracts from the yeast diploid strain ccdYK or from the yeast haploid strains cdYK×pyHKm and cdYK×pyHKc, lacking the yeast KRS1 gene but complemented with human mito-KRS or cyto-KRS, respectively, were analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and stained with Coomassie blue (left panel) or subjected to Western blotting using antibodies directed to yeast LysRS (IgG × yKRS) or raised against the full-length cytoplasmic LysRS of mammalian origin (IgG × KRS). The polypeptides corresponding to mito-LysRS and cyto-LysRS, visualized by Coomassie blue staining, are indicated by dots. MARS, the multi-aminoacyl-tRNA synthetase complex purified from rabbit containing cyto-LysRS (indicated by an arrow). *, a polypeptide cross-reacting with anti-yKRS antibodies.