FIG. 6.

Subcellular localization of luciferase in oleate-grown cells of S. cerevisiae transformed with the luciferase-SKL construct expressed under control of the CTA1 promoter (see Materials and Methods). (A) Luciferase-expressing wild-type cells were fractionated by differential centrifugation of a homogenate (H) into a 17,000 × g pellet (P) and supernatant (S), followed by the measurement of 3-hydroxyacyl-CoA dehydrogenase (3HAD) and luciferase activity. (B) The 17,000 × g pellet (P) was further fractionated by Nycodenz equilibrium density gradient centrifugation (fractions 1 to 12). Mitochondrial (M) and peroxisomal (P) matrix markers are fumarase (▴) and 3-hydroxyacyl-CoA dehydrogenase (3HAD) (●), respectively. Luciferase activity (solid bars) was measured in the fractions (see Materials and Methods).