1887

Abstract

The three plus-strand genomic RNAs of (AMV) and the subgenomic messenger for viral coat protein (CP) contain a 5′-cap structure, but no 3′-poly(A) tail. Binding of CP to the 3′ end of AMV RNAs is required for efficient translation of the viral RNAs and to initiate infection in plant cells. To study the role of CP in translation, plant protoplasts were transfected with luciferase (Luc) transcripts with 3′-terminal sequences consisting of the 3′ untranslated region of AMV RNA 3 (Luc–AMV), a poly(A) tail of 50 residues [Luc–poly(A)] or a short vector-derived sequence (Luc–control). Pre-incubation of the transcripts with CP had no effect on Luc expression from Luc–poly(A) or Luc–control, but strongly stimulated Luc expression from Luc–AMV. From time-course experiments, it was calculated that CP binding increased the half-life of Luc–AMV by 20 % and enhanced its translational efficiency by about 40-fold. In addition to the 3′ AMV sequence, the cap structure was required for CP-mediated stimulation of Luc–AMV translation. Glutathione -transferase pull-down assays revealed an interaction between AMV CP and initiation factor complexes eIF4F and eIFiso4F from wheatgerm. Far-Western blotting revealed that this binding occurred through an interaction of CP with the eIF4G and eIFiso4G subunits of eIF4F and eIFiso4F, respectively. The results support the hypothesis that the role of CP in translation of viral RNAs mimics the role of the poly(A)-binding protein in translation of cellular mRNAs.

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2005-06-01
2024-12-08
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