The eukaryotic mRNA 3′ poly(A) tail and the 5′ cap cooperate to synergistically enhance translation. This interaction is mediated, at least in part, by eIF4G, which bridges the mRNA termini by simultaneous binding the poly(A)-binding protein (PABP) and the cap-binding protein, eIF4E. The poly(A) tail also stimulates translation from the internal ribosome binding sites (IRES) of a number of picornaviruses. eIF4G is likely to mediate this translational stimulation through its direct interaction with the IRES. Here, we support this hypothesis by cleaving eIF4G to separate the PABP-binding site from the portion that promotes internal initiation. eIF4G cleavage abrogates the stimulatory effect of poly(A) tail on translation. In addition, translation in extracts in which eIF4G is cleaved is resistant to inhibition by the PABP-binding protein 2 (Paip2). The eIF4G cleavage-induced loss of the stimulatory effect of poly(A) on translation was mimicked by the addition of the C-terminal portion of eIF4G. Thus, PABP stimulates picornavirus translation through its interaction with eIF4G.