Mutations that increase the activity of a transcriptional activator in yeast and mammalian cells

Activating region I of GAL4 protein is a stretch of amino acids, positioned adjacent to the DNA-binding region, that activates transcription in yeast and, as we show here, in mammalian cells. Here we describe mutations located throughout a 65-amino acid region that increase the activation function of region I. Most of these mutations replace positively charged amino acids in the region with neutral ones, although we also describe substitutions at one position that do not alter the charge of the region. Mutations of region I that alter the activation function in yeast have similar effects on activation when assayed in mammalian cells. When individual mutations that raise the acidity of the activating region are recombined, the activities of the mutant proteins increase with increasing negative charge in both yeast and mammalian cells. These results extend and modify the correlation between acidity and activation and suggest that the requirements for a strong activating region are conserved in yeast and mammals.