Two Distinct Domains in Staf To Selectively Activate Small Nuclear RNA-Type and mRNA Promoters

ABSTRACTStaf is a transcriptional activator of prime importance for enhanced transcription of small nuclear (snRNA) and snRNA-type genes transcribed by RNA polymerases II and III (Pol II and III). In addition to this activity, it also possesses the capacity to stimulate expression from an RNA polymerase II mRNA promoter. This promiscuous activator thus provides a useful model system for studying the mechanism by which one single transcription factor can activate a large variety of promoters. Here, we report the use of in vivo assays to identify the Staf activation domains involved in promoter selectivity. Analysis of Staf mutants reveals the existence of two physically and functionally distinct regions, outside of the DNA binding domain, responsible for mediating selective transcriptional activation. While a 93-amino-acid domain, with the striking presence of four repeated units, is specialized for transcriptional activation of an mRNA promoter, a segment of only 18 amino acids, with a critical Leu-213 residue, acts specifically on Pol II and Pol III snRNA and snRNA-type promoters. In addition, this study disclosed the fundamental importance of invariant leucine and aspartic acid residues located in each repeat unit of the mRNA activation domain. Staf is therefore the first transcriptional activator described so far to harbor two physically and functionally distinct activator domains. This finding suggests that the same activator can contact different, specialized transcription complexes formed on different types of basal promoters through promoter-specific transactivation pathways. ACKNOWLEDGMENTSWe are grateful to E. Myslinski for critical reading of the manuscript. We thank J. Hoffmann and C. Kappler forDrosophila cell cultures, P. Remy for microinjection facilities, A. Hoeft for oligonucleotide synthesis, and C. Loegler for excellent technical assistance.This work was supported by grants from the Université Louis Pasteur in Strasbourg, the Association pour la Recherche sur le Cancer (ARC) and the European Union (EEC Biotech Program B102-CT92-0090).