The GAL11 gene encodes an auxiliary transcription factor required for full expression of many, if not all, genes of the yeast Saccharomyces cerevisiae. We have recently shown that GAL11-encoded protein (Gal11p) enhances basal transcription from the CYC1 promoter in a cell-free transcription system [(1993) Proc. Nail. Acad. Sci. USA 90, 8382-8386]. Here we indicate that Gal11p stimulates basal transcription in a gene-specific manner in vitro. We further suggest that the mechanism underlying the transcriptional stimulation by Gal11p is distinct from that by DNA-bound activators, since Gal11p stimulated transcription in a reaction system where activators were unable to enhance transcription due to the lack of intermediary factors.