FIG. 8.

Time course analysis of the physical alterations to Ssy1p, Ptr3p, and Ssy5p after induction of sensor function by l-leucine. Strains HKY20, HKY33, and HKY77 expressing SSY1-HA1, PTR3-HA1, and SSY5-c-myc, respectively, were grown in liquid cultures of SD to an OD₆₀₀ of 0.5 (t = 0; lane 1). At t = 0, each culture received an aliquot of l-leucine to a final concentration of 1.3 mM (lanes 2 to 6). At the times indicated, subsamples were removed, whole-cell lysates were prepared, and proteins were analyzed by immunoblotting (upper panel). Lane 7 shows protein levels present in an uninduced control culture similarly incubated for 180 min. The corresponding chemiluminescent signals were quantitated (lower panel).