The cis-acting gibberellin response complex in high-pI alpha-amylase gene promoters. Requirement of a coupling element for high-level transcription

In cereal alpha-amylase gene promoters the cis-acting gibberellin response element (GARE) is required for increased transcription in the presence of gibberellin. In low-isoelectric point (pI) alpha-amylase gene promoters a second type of cis element, termed a coupling element, must also be present in a specific position near the GARE; otherwise, the level of transcription in the presence of gibberellin is only a few percent of maximum. The coupling element may help determine where and when in development high-level, hormonally regulated transcription will occur. Such coupling elements have not yet been shown to be necessary for high-level transcription from high-pI alpha-amylase gene promoters. Here we use quantitative transient expression assays to show that a high-pI promoter truncated to -300 is a weak promoter due to the absence of a functional coupling element in the vicinity of the GARE. Gibberellin-induced transcription increases substantially when coupling element function is provided, either by appending upstream regions normally attached to the promoter or by inserting a defined coupling element from a low-pI promoter. Thus, in a second type of gibberellin-regulated promoter coupling element function was found to be crucial for hormone regulation to result in high-level transcription