Fig. 4. Yeast two-hybrid survey of the sites in myotilin mediating interaction with FATZ-1. The PJ69-4A yeast strain was co-transformed with pACT2 plasmids encoding various deletion mutants of myotilin in fusion with GAL4-activation domain (AD) and pAS2-1 plasmids encoding FATZ-1 fused at its N-terminus with GAL4-DNA-binding domain (BD). The N-terminal serine-rich domain and the two Ig-like domains of myotilin are represented by a dark grey box and the black loops, respectively. Growth was estimated after 5 to 10 days of incubation at 30°C. +, growth; –, no growth, on selective media. The results indicate an interaction between myotilin^1-498, myotilin^S55F or myotilin^T57I and FATZ-1, whereas shorter myotilin constructs did not interact. Transactivation controls were systematically performed for each construct with the opposite vectors without insert. Each experiment was repeated at least twice.