Figure 8.

Trans-Activation of pH4A748-, Mutated pH4A748-, and Oct Motif-Controlled Reporters by Trans-Activation Domain-Fused NtWLIM2. (A) Reporter and effector constructs used in this study. H4 reporter was as previously described (Figure 4). The 2xOCT reporter plasmid contained two tandemly oriented copies of the EMSA oligonucleotide 2xOCT fused upstream to the –46CaMV 35S core promoter (35S core) that controlled LUC expression. In mtH4, LUC was under the control of a mutated version of pH4A748 that comprised point mutations within the octamer and degenerated octamer elements. The effector construct AD–NtWLIM2 consisted in a fusion of the trans-activation domain (AD) of herpes simplex virus protein (Sadowski et al., 1988) to NtWLIM2. An unfused AD construct was used as a negative control effector. (B) Level of trans-activation of pH4A748-, mutated pH4A748-, and 2xOCT motif-controlled reporters by AD-fused NtWLIM2 in Arabidopsis protoplasts. The LUC activity measured in the presence of 1μg of effector plasmid is given relative to the LUC activity measured in the presence of 1μg of GFP-expressing vector, the latter being set to 100% (white bars, control). The two used effector constructs allowed ectopic expression of either AD–NtWLIM2 (black bars) or unfused AD (dark gray bars; AD). Data represent mean values of three independent transfections and error bars indicate standard deviations.