Genetic isolation of ADA2: a potential transcriptional adaptor required for function of certain acidic activation domains
Department of Medicine, Division of Infectious Diseases and Immunology
Amino Acid Sequence; Base Sequence; DNA Mutational Analysis; *Genes, Fungal; Genome; Molecular Sequence Data; Saccharomyces cerevisiae; Sequence Alignment; *Trans-Activators
Immunology and Infectious Disease
We have devised a genetic strategy to isolate the target of acidic activation domains of transcriptional activators based on toxicity in yeast cells of the chimeric activator, GAL4-VP16. Toxicity required the integrity of both the VP16 acidic activation domain and the GAL4 DNA-binding domain, suggesting that inhibition resulted from trapping of general transcription factors at genomic sites. Mutations that break the interaction between GAL4-VP16 and general factors would alleviate toxicity and identify transcriptional adaptors, if adaptors bridged the interaction between activators and general factors. We thus identified ADA1, ADA2, and ADA3. Mutations in ADA2 reduced the activity of GAL4-VP16 and GCN4 in vivo. ada2 mutant extracts exhibited normal basal transcription, but were defective in responding to GAL4-VP16, GCN4, or the dA:dT activator. Strikingly, the mutant extract responded like wild type to GAL4-HAP4. We conclude that ADA2 potentiates the activity of one class of acidic activation domain but not a second class.
DOI of Published Version
Cell. 1992 Jul 24;70(2):251-65. Link to article on publisher's site
Berger, Shelley L.; Pina, Benjamin; Silverman, Neal S.; Marcus, Gregory A.; Agapite, Julie; Regier, Jeffrey L.; Triezenberg, Steven J.; and Guarente, Leonard, "Genetic isolation of ADA2: a potential transcriptional adaptor required for function of certain acidic activation domains" (1992). Infectious Diseases and Immunology Publications and Presentations. 29.