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Figure 1


Fig. 1. Identification of proteins that interact with Ste20. (A) The split-ubiquitin system. See text for details. Degraded RUra3 is shown on the right. X represents any interactor of Ste20. (B) Ste20 interacts with Erg4, Ncp1 and Cbr1 in vivo using the split-ubiquitin technique. 105 cells of the indicated plasmid combinations were spotted onto media either lacking histidine and leucine to select for the plasmids or lacking histidine, leucine and uracil to monitor protein interactions. The unrelated proteins Rdi1 and Ubc6 served as negative controls. Bem1 is known to interact with Ste20. (C) Ste20 interacts with Erg4 in vitro. Purified GST and GST-Ste20 that were bound to glutathione-sepharose beads were incubated with a yeast lysate of ERG4-9MYC cells. Eluted proteins were analyzed by immunoblotting using anti-Myc antibodies. (D) Ste20 interacts with Cbr1. The experiment was performed as in C using a yeast lysate of 3HA-CBR1 cells. Proteins were detected with anti-HA antibodies.





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