Figure 3.

DAF-16 interacts with p300 and SRC. (A) Interaction of [³⁵S]methionine-labeled DAF-16 with the KIX and C/H3 interaction domains of GST⋅CBP. In vitro translated [³⁵S]methionine-labeled DAF-16 or DAF-16 (Δ340–511) was incubated with GST (lane 1); or GST⋅CBP (KIX), which encodes amino acids 450–684 of CBP (lane 2); or GST⋅CBP (C/H3), which encodes amino acids 1890–2441 of CBP (lane 3); or GST⋅SRC, which encodes amino acids 594–780 (lane 4). The bound proteins were washed, eluted, and subjected to SDS/PAGE as described in the text. The autoradiograph of a dried gel is shown. (B) Interaction of GST⋅SRC with [³⁵S]methionine-labeled p300 and DAF-16. In vitro translated [³⁵S]methionine-labeled p300 (lanes 2 and 4) or DAF-16 (lanes 1 and 3) was incubated with bacterially produced GST (lanes 1 and 2) or GST⋅SRC (amino acids 594–780) (lanes 3 and 4) bound to glutathione-Sepharose beads. Eluted proteins are shown. (C) Interaction of [³⁵S]methionine-labeled DAF-16, FKHR, and AFX with GST⋅CBP (KIX) and GST⋅SRC. [³⁵S]Methionine-labeled DAF-16, FKHR, and AFX were incubated with GST (lane 1), GST⋅KIX (lane 2), or GST⋅SRC (lane 3) bound to glutathione-Sepharose beads. Eluted proteins are shown.