FIG. 1.

Interaction of Tim44 and the ATPase domain of mtHsp70 (Ssc1) in the two-hybrid system. (A) Two-hybrid interaction of Ssc1(APC) and Ssc1(A) with Tim44. The reporter yeast strain Y190 was transformed with the two-hybrid plasmids expressing the indicated fusion proteins. Transformants were grown for 2 days at 30°C on minimal medium lacking tryptophan and leucine. One clone of each transformation was streaked out on minimal medium lacking tryptophan, leucine, and histidine but containing 3-aminotriazole and cultured for 3 days at 30°C. (B) The two-hybrid fusion proteins are expressed in yeast. The yeast strain Y190 was transformed as described in the legend to panel in A. One clone of each transformation was cultured overnight in minimal medium lacking tryptophane and leucine. Cells of a 1-ml liquid culture were resuspended in 15 μl of electrophoresis sample buffer and separated by SDS-PAGE. The proteins were detected by immunodecoration with antibodies directed against Hsp70 or Tim44. endog., endogenous (authentic) Hsp70s and Tim44 reacting with the antibodies.