FIGURE 5:

TPRs are not required for stable association with cargo. (A) Schematic representation of the interaction between the cargo, Chs3p, and the exomer–cargo recognition subunit, Chs6p. (B) Chromosomally generated Chs6p TPR mutants. (C) Chs6(ΔTPR5) and Chs6(LG-WD) interact with cargo. To assess cargo interaction, Chs3p-2GFP was precipitated from DSP cross-linked lysates with anti-GFP monoclonal antibodies, and the precipitates were probed for different Chs6p constructs. Control immunoprecipitations were performed using monoclonal HA antibody. Chs6(ΔTPR5) and a double TPR5 point mutant Chs6(LG-WD) retained association with Chs3p. (D) Chs6(ΔTPR1-4) interacts with Chs3p and does so independently of Chs5p, the core exomer subunit. The cross-linker immunoprecipitation was performed as described in B.