Figure 1
YBAP1 interacts with the C1 and C2 regions of the YB-1 tail domain. (a) HeLa cell lysates were treated with (lanes 4 and 6) or without (lanes 3 and 5) RNase A, and subjected to immunoprecipitation with control rabbit IgG (lanes 3 and 4) or rabbit anti-YB-1 antibodies (lanes 5 and 6). Immunoprecipitates and cell lysates (10% of input, lanes 1; 2.5% of input, lane 2) were analyzed by immunoblotting with antibodies against YB-1, YBAP1, and PABP. (b) FLAG-tagged YB-1 and its fragments were synthesized in wheat germ extracts and incubated with GST (lanes 7, 9, 11, 13, 15, and 17) or GST-YBAP1 (lanes 8, 10, 12, 14, 16, and 18). Protein complexes were bound to glutathione–Sepharose, and the eluted materials together with the input extracts (lanes 1–6) were analyzed by immunoblotting with anti-FLAG antibodies. Schematic diagrams of the YB-1 fragments are shown at the bottom. +++ and −−− indicate basic and acidic/aromatic amino acid clusters, respectively.
