Figure 2

IR induced disassembly of the eIF4G1-dependent as well as DAP5-dependent initiation complex. Jurkat cells were irradiated with 10 Gy. (A) Lysates were made 4–24 h after IR. Cleavage of DAP5 was detected already 8 h after irradiation. (B) Jurkat cells were irradiated with 10 Gy and treated with 30 μM of caspase inhbitor zVAD or the respective amount of solvent. 24 h after irradiation, cells were lysed. Treatment with zVAD abrogated IR-induced cleavage of DAP5. (C) 24 h after IR with 10 Gy, cells were lysed and a pull-down assay was made using 7-methyl GTP agarose. DAP5 was not detected in the cap-dependent initiation complex. 24 h after IR with 10 Gy, cells were lysed and DAP5 (D), and eIF4G1 (E) were precipitated. (D) Less eIF4A and eIF3A co-precipitated with DAP5 (D) and eIF4G1 (E) after IR indicating disassembly of the DAP5-dependent and eIF4G1-dependent initiation complexes.