Figure 6.

Fus2p and Kel1p interact in pheromone-induced cells. (A) Wild-type Fus2p interacts with Kel1p. KEL1 tagged with 3× HA was either integrated (Int) at the KEL1 locus (MY15063) or cloned on to a 2μ vector (pMR6953). These constructs along with untagged KEL1 (−) were pulled down with anti-HA magnetic beads. Interaction with GFP-tagged Fus2p (pMR5482) was assessed via Western blot with anti-GFP antibodies. (B) All Fus2p C-terminal truncations interact with Kel1p. Co-immunoprecipitations were performed as in A with strains containing Fus2p^1-640 (pMR6854), Fus2p^1-650 (pMR6853), Fus2p^1-660 (pMR6854), or Fus2p^1-670 (pMR6775). (C and D) Kel1p has two binding sites on Fus2p. (C) Interaction with Kel1p was tested with strains containing Fus2p^105-677 (pMR5784), Fus2p^1-580 (pMR5886), Fus2p^∆105-415 (pMR5883), Fus2p^1-415 (pMR7008), and (Fus2p^1-104 (pMR5774). Because the Fus2p^1-104 fragment is much smaller than wild-type Fus2p (38 kDa vs. 102 kDa), we show the input Fus2p and bound Fus2p panels with the center removed, denoted by a black line. The asterisk denotes where Fus2p^1-104 would run if it bound Kel1p-HA. (D) Map of all Fus2p fragments tested summarizing the results of the binding experiments.