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. 1998 Jun 1;12(11):1665–1677. doi: 10.1101/gad.12.11.1665

Figure 4.

Figure 4

Figure 4

 ATP prevents RNA from competing with eRF3 for binding to Upf1p. (A) Reaction mixtures were prepared as described in Fig. 3A, except that binding was performed in IBTB, and reaction mixtures contained 1 mm ATP or poly(U) RNA at the concentrations indicated above each lane. The reaction mixtures were mixed for 1 hr at 4°C. Following mixing, the complexes were washed with IBTB containing 1 mm ATP or poly(U) RNA at concentrations as indicated above each lane. (B) Upf1pK436A interacts weakly with eRF1. Reaction mixtures were prepared as in Fig. 1B, substituting Upf1pK436A for the wild-type protein (lanes 5–8). (C) A mutant Upf1p is unable to interact with eRF3 in the presence of RNA. Reaction mixtures were prepared as in A, substituting purified Upf1pK436A for the wild-type protein. Reactions contained 1 mm ATP or 40 μg/ml poly(U) RNA as indicated above each lane.