Ribosome shunting in the cauliflower mosaic virus 35S RNA leader is a special case of reinitiation of translation functioning in plant and animal systems

The shunt model predicts that small ORFs (sORFs) within the cauliflower mos aic virus (CaMV) 35S RNA leader and downstream ORF VII are translated by di fferent mechanisms, that is, scanning-reinitiation and shunting, respective ly. Wheat germ extract (WGE) and rabbit reticulocyte lysate (RRL) in vitro translation systems were used to discriminate between these two processes a nd to study the mechanism of ribosomal shunt. In both systems, expression d ownstream of the leader occurred via ribosomal shunt under the control of a stable stem and a small ORF preceding it. Shunting ribosomes were also abl e to initiate quite efficiently at non-AUG start codons just downstream of the shunt landing site in WGE but not in RRL. The short sORF MAGDIS from th e mammalian AdoMetDC RNA, which conditionally suppresses reinitiation at a downstream ORF, prevented shunting if placed at the position of sORF A, the 5'-proximal ORF of the CaMV leader. We have demonstrated directly that sOR F A is translated and that proper termination of translation at the 5'-prox imal ORI: is absolutely required for both shunting and linear ribosome migr ation. These findings strongly indicate that shunting is a special case of reinitiation.