FIGURE 3:

Arg5 and Arg6 can be imported separately in vitro and in vivo. (A-C) Radiolabeled precursor proteins of Arg6^1–502, Arg5^503–862, and Su9-Arg5^503–862 were incubated with isolated mitochondria for the indicated times and analyzed by SDS–PAGE and autoradiography. Nonimported material was digested with proteinase K (left half). Twenty percent of the total lysate used per import lane is loaded for control. The membrane potential (Δψ) was depleted with VAO. Red arrowheads indicate processing sites. p, precursor, m, mature. (D, E) Yeast cells that lack endogenous Arg5,6 (Δarg5,6) were transformed with plasmids for expression of the indicated Arg5,6 variants and streaked out on plates containing minimal growth medium without arginine.