Viral tRNA-like structure hijacks host ribosomes for poly(A)-independent translation

Positive-sense RNA viruses often use 3′ tRNA-like structures (TLSs) instead of poly(A) tails to capture the host translation machinery. While TLSs resemble canonical tRNAs and engage host factors, their ability to directly recruit ribosomes has remained unresolved. Here, we present cryo-electron microscopy snapshots of the histidine-accepting TLS (TLS ^His ) from tobacco mosaic virus bound to the 60 S subunit, the 80 S ribosome, and the 80 S -tRNA _i ^Met initiation complex. Across these states, TLS ^His is consistently anchored to the L1 stalk of 60 S , even under cycloheximide treatment, yet remains dynamic on the 40 S subunit. Structural analysis shows that TLS ^His transitions from the E-site to an adjacent Z-site to accommodate initiator tRNA. Functional assays show that TLS ^His preferentially associates with ribosomal subunits over polysomes and can substitute for a poly(A) tail to promote robust cap-dependent translation. Together, these findings reveal how a viral tRNA mimic hijacks host ribosomes to promote poly(A)-independent translation.