PMC

Sl-eIF4E1 G1485A mutant and the corresponding Hm-WT were inoculated with PVY-LYE90, PVY-LYE84, TEV-HAT or PepMoV-Texas. (A) At 15 dpi, plants were assayed for potyviral coat protein accumulation by DAS-ELISA in non-inoculated leaves. (B) PVY-LYE90 and PepMoV RNA accumulation was assessed by RT-PCR in inoculated (il) and systemic leaves (sl). Mock indicates non inoculated M82 plants.

Black boxes represent the exons. Lanes linking exons indicate introns. Dashed lines indicate the genomic regions screened for mutations. Triangles pointing up indicate mutations in coding regions, whereas those pointing down indicate mutations in noncoding regions. Red, black and grey triangles represent alterations causing truncations, missense and silent mutations, respectively. Only exons 7 to 9 are shown for Sl-eIF4G.

(A) Total RNA was extracted from leaf tissues of the Hm-WT and G1485A mutant and full lengh cDNAs were amplified and analysed on 1% agarose gel. (B) Northern analysis of Sl-eIF4E1 from Hm-WT and G1485A mutant lines, using exon 1 as probe. (C) Representation of the cDNA structure of the wildtype and the G1485A mutant form. The G1485A mutation is shown by a dashed rectangle.

(A) Western blot analysis of total soluble leaf protein of N. tabacum, S. lycopersicum Hm-WT and Sl-eIF4E1 G1485A mutant probed with an antibody raised against N. tabacum Nt-eIF4E1. (B) Soluble protein extracts of the Hm-WT and G1485A mutant were purified by affinity chromatography on m7G-sepharose column. Total protein extract (lane 1), the flow through (lane 2), the wash (lane 3) and the bound eIF4E proteins eluted with an m7GDP-cap analogue were analysed by Western blot, using Nt-eIF4E antibody.