FIG. 2.
The N terminus of Cox2(1-109)-HA is translocated across the inner membrane immediately after synthesis. Log-phase cells grown in synthetic complete medium containing raffinose and lacking methionine were treated with cycloheximide for 5 min prior to incubation with [35S]methionine for 10 min. Crude mitochondria were isolated, electrophoresed on a 14 to 20% gradient SDS gel, and visualized by autoradiography. All strains harbored the cox2(1-109)-HA mitochondrial construct. The relevant nuclear genotypes of strains HLF3-60, HLF4-73, HLF4-17, HLF4-30, and HLF4-63 are indicated. N-terminally processed (filled arrow) and unprocessed (unfilled arrow) Cox2(1-109)-HA peptides are indicated.