Figure 2. Import of tRK1 into yeast (Fig. 2A), rat liver (Fig. 2B) and human mitochondria (Fig. 2C) in the presence of human mitochondrial lysyl-tRNA synthetase.

Import of tRK1 into isolated yeast or mammalian mitochondria was performed in the presence or absence (lane 2) of either yeast or human mitochondrial lysyl-tRNA synthetase (lanes 3–7). The import efficiency was assessed by RNase protection assay followed by polyacrylamide gel electrophoresis and phosphorimaging. Lane 1 contains wild type yeast cytosol that presumably contains both cytosolic and mitochondrial lysyl- tRNA synthetase. Fig. 2D. Substitution of yeast cytosol with human cytosol for the import of tRK1 into either yeast or mammalian mitochondria. Import of tRK1 was carried out into yeast (lanes 1, 2, 4 & 9) and human mitochondria (lanes 3, 5, 6, 7 & 8) in the presence of wild type yeast cytosol (lanes 2 & 3), or mskΔ cytosol (lanes 4, 7 & 8) or human cytosol (lanes 5, 8 & 9). The import efficiency was assessed by RNase protection assay followed by electrophoresis. The bands were quantified and relative tRK1 import values were mentioned by taking maximum imported sample as a value of 1.