We have combined in vivo genomic footprinting and light-induced transient expression of chalcone synthase promoter derivatives in parsley protoplasts to identify cis sequences regulating light activation. The parsley chalcone synthase promoter contains two cis "units" that are light-responsive. Each unit is composed of short DNA stretches of approximately 50 base pairs, and each contains two in vivo footprints. One of the footprints in each unit covers a sequence that is highly conserved among other light- and stress-regulated plant genes. The other footprinted sequences in each unit are not related to each other. The TATA distal light-responsive unit is inherently weak but can compensate partially for the loss of the stronger TATA proximal unit. Levels of light-induced expression from either can be influenced by the presence of a region of approximately 100 base pairs located upstream of the TATA distal light-responsive unit. Combination of the light-responsive units and upstream region generates a synergistic response to light. We speculate that functional compensation generated by nonidentical, but sequence-related, cis units foreshadows combinatorial diversity of cognate trans factors.
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