FIG. 3.

Interaction of AR AF-1 with C-terminal regions of p160 coactivators in vitro. Glutathione-Sepharose-bound GST or GST-ARAF1 was incubated with ³⁵S-labeled full-length GRIP1, SRC-1a, SRC-1e, or their fragments translated in vitro from pSG5.HA vectors. Bound proteins were eluted and analyzed by SDS-PAGE and autoradiography; shown for comparison is 10% of the total labeled protein incubated in each binding reaction (10% input). Functional domains in diagrams of the coactivator fragments are represented as in Fig. 1A. Coactivator fragments translated in vitro include the following amino acids: GRIP1 5 to 1462 (full length), 5 to 1121, 1122 to 1462, and 1305 to 1462; SRC-1a 1 to 1441 (full length), 1 to 977, 977 to 1441, and 1172 to 1441; SRC-1e 1 to 1399 (full length), 1 to 977, 977 to 1399, and 1172 to 1399.