Figure 1

The closed loop complex and 4E-BP repression complexes are maintained during purification. (A) A diagram depicting the closed loop complex with eIF4E (4E), eIF4G (4G) and PABP bound to the mRNA; the 4E-BPs Caf20p and Eap1p are also represented competing with the eIF4E-eIF4G interaction. (B) Western blots probed with a protein A peroxidase (PAP) conjugate which detects the TAP-tagged proteins labeled above the blots. Input, flowthrough and eluates are presented on the top, middle and bottom blots respectively. The vast majority of TAP-tagged proteins purified appear in the eluates. (C) Western blots probed with PAP or the antibodies depicted on the right, which detect the components of the closed loop complex (either TAP-tagged or not) depicted on the left. Samples are eluates from TAP-affinity chromatography using strains bearing the TAP-tagged protein depicted above each lane. (D) As for (C), except the components of the 4E-BP complexes were assessed by TAP affinity chromatography.