FIGURE 7.

Role of Ser/Arg-rich motif in the interaction of DhPpz1p with DhHal3p. A, two-hybrid assay for interactions between DhHal3p and DhPpz1p. Wild type DhPPZ1 or its mutant having a deletions in the Ser/Arg motif (Δ27–36) cloned in pEG202 vector was used as bait, whereas DhHAL3 cloned in plasmid pJG4-5 was used as prey. Bait and prey constructs (in pairs as indicated) or in combination with empty prey or bait (as control) were transformed into S. cerevisiae strain EGY48. Growth of the transformants on Gal-Raf minimal medium is shown after dilution spotting. Experiments were repeated twice with pool of four independent transformants. Representative data are shown. B, growth of EGY48 harboring DhPPZ1 and Δ27–36 bait along with empty prey or DhHal3 in prey vector were grown overnight in minimal media with 2% raffinose (without tryptophan and histidine). The cultures were re-inoculated in minimal media with 1% raffinose and 2% galactose (without tryptophan, histidine, and leucine) at A₆₀₀ ∼0.10 and grown further for 41 h. Data (mean ± S.D.) of two independent experiments, each performed in duplicate with a pool of four different transformants, is shown here.