Wild-type and mutant Mss116 proteins were expressed at similar levels in yeast. Immunoblot of Flag-tagged wild-type (WT) Mss116p, Mss116p lacking the mitochondrial targeting sequence (Δmts), Mss116p lacking the mitochondrial targeting sequence with an inactivating mutation in the DEAD motif (ΔmtsE268Q), and Mss116p lacking the mitochondrial targeting sequence with an inactivating mutation in the P-loop (ΔmtsK158A). Coexpression of chimeric self-cleaving mRNA (HP44) with Mss116p (Δmts) did not affect Mss116p expression. After normalization using PSTAIRE protein loading controls, results obtained with different samples of the same yeast culture and with different yeast cultures varied less than twofold.
