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. 2006 Jan 30;103(6):1988–1993. doi: 10.1073/pnas.0505667103

Fig. 3.

Fig. 3.

Transient transactivation analysis of amino acid-substituted AREB1. Transient transactivation of the RD29B promoter-GUS fusion gene by AREB1 and its amino acid-substituted proteins by using Arabidopsis protoplasts prepared from T87 suspension cells. The reporter GUS gene driven by the 77-bp DNA fragment of the RD29B promoter containing two ABREs was transfected into T87 protoplasts with each effector plasmid or the vector plasmid (pBI35SΩ) as a control. To normalize for transfection efficiency, the CaMV 35S promoter/luciferase plasmid was cotransfected in each experiment. Each effector plasmid contains the CaMV 35S promoter and TMV Ω sequence fused to wild-type or amino acid-substituted AREB1 ORF. Bars indicate the SD of triplicates. Transformed protoplasts were incubated with (open) or without (solid) 50 μM ABA in culture medium, at 22°C for 16–20 h in the dark in all transient transactivation experiments shown. (A) Effects of each single substitution of the R-X-X-S/T sites. (B) Effects of multiple amino acid substitutions (Ser/Thr to Ala). M1; S94A, M2; S(86, 94)A, M3; S(26, 86, 94)A, M4; S(26, 86, 94)A and T135A, M5; S(26, 86, 94, 413)A and T135A. (C) Effects of single or multiple amino acid substitutions (Ser/Thr to Asp). M6; S94D, M7; S(26, 86, 94, 413)D, M8; S(26, 86, 94, 413)D and T135D.