Figure 3.

Interaction between various recombinant TnT, TnI, and TnC polypeptides as measured by percent RPE (RPE%) in the yeast two hybrid system. RPE% was calculated as described in the Materials and Methods. Bars = SD. (a) Interaction of wild-type and mutant TnT proteins with TnI. Wild-type TnI (TnI-wt) was coexpressed with wild-type TnT (TnT-wt) or the mutant TnTs (TnT-G²⁰³S²⁰⁴, TnTΔ203–214 and TnT-R²⁰⁷) in yeast, and the activation of the LEU2 reporter gene was measured as RPE%. A wild-type TnT coexpressed with pJG4–5, the transcriptional activator alone was included as a negative control (TnT-wt +pJG4–5). (b) Interaction of the truncated TnI_58–108 fragment containing the HR domain with various TnT constructs. TnI_58–108 was coexpressed with either TnT_150–258, or TnT-wt, or TnTΔ203–214, or TnT-G²⁰³S²⁰⁴, or TnT-R²⁰⁷. As a control, TnI_58–108 was also coexpressed with the bacterial LexA repressor DNA binding domain alone (pEG202). (c) Interaction of TnC with wild-type and mutant TnT proteins. TnC was coexpressed with wild-type TnT (TnT-wt) or the mutant TnTs (TnT-G²⁰³S²⁰⁴ and TnTΔ203–214), and the activation of the LEU2 reporter gene was measured as RPE%. A wild-type TnT expressed without any interactor protein is included as a negative control (TnT-wt alone).