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. 2014 Jul 10;42(15):10005–10022. doi: 10.1093/nar/gku584

Figure 2.

Figure 2.

Cap-associated factors were genetically linked to Ded1. Cells transformed with plasmids expressing the indicated proteins were serially diluted by factors of 10, spotted on synthetic minimal-medium plates lacking tryptophan (p424 plasmids) or uracil (p426 plasmids) and incubated at the indicated temperatures. Plates were incubated for 3 days at 30 and 36°C, and for 8 days at 18°C. (A) Cells expressing the slow-growth mutant Ded1-F162C of the Q motif were transformed with plasmids overexpressing the indicated proteins. The p424 was an empty plasmid control. (B) Pab1 and eIF4G1 were inhibitory when overexpressed in cells expressing wildtype Ded1. (C) Multicopy suppression of cells expressing the slow-growth mutant Ded1-F405C of motif IV. Cells were transformed with plasmids overexpressing Cbp20 and Cbp80. (D) Ded1 was a multicopy suppressor of cells deleted for cbp20. (E) Cells expressing Ded1 mutated in the eIF4E-binding motif (Ded1-4E; Y21A/L26A) gave a slow-growth phenotype relative to the wild type (Ded1) when expressed off the low copy p415-PL plasmid.