FIGURE 3.

DdiTLP1 catalyzes Thg1 activity in vivo in yeast. Complementation of the yeast thg1Δ growth defect by each of the D. discoideum TLPs was tested using a plasmid shuffle assay, as previously described (Abad et al. 2010). DdiTLPs, as indicated, expressed under control of a galactose-inducible promoter on LEU2 plasmids, were transformed into the yeast thg1Δ strain and two independent positive transformants were tested by replica plating onto the indicated media. Clones of TLP2 and TLP3 lacked their respective mitochondrial targeting sequences and the Q/N-rich sequence for TLP3 (see Fig. 1). Empty vector control strains (vect) were also tested. Photographs were taken after 3–4 d growth at 30°C.