title: Activating regions of yeast transcription factors must have both acidic and hydrophobic amino acids.
abstract: Two similarities among transcriptional activating regions of many eukaryotic transcription factors, like those from GAL4, GCN4, and VP16, are that they have a net negative charge, and that many of them can potentially form amphipathic alpha-helices with acidic amino acids on the hydrophilic face. Based on these similarities, E. Giniger and M. Ptashne previously designed a short peptide (AH) which is predicted to have the potential to form a negatively charged amphipathic alpha-helix; AH was able to mediate transcription activation in yeast when it was attached to the DNA binding and dimerization portion of GAL4 [GAL4(1-147)]. This paper describes screening of a pool of AH derivatives containing randomized amino acids fused to GAL4(1-147) and to an analogous region of LexA [LexA(1-87)] in yeast strains. Results suggest that both acidic and hydrophobic amino acids are critical features of activating regions--these results are consistent with the model that activating regions often form amphipathic alpha-helices. This work is novel because hydrophobic amino acids are also shown to be important in activating regions of yeast transcription factors.
authors.affiliation: Department of Biochemistry and Molecular Biology, Harvard University, Cambridge, MA 02138.
authors.forename: D M
authors.initials: DM
authors.lastname: Ruden
mesh_terms: D000595:Amino Acid Sequence; D000596:Amino Acids / Q000378:metabolism*; D001426:Bacterial Proteins / Q000378:metabolism; D001483:Base Sequence; D004271:DNA, Fungal; D004268:DNA-Binding Proteins; D005656:Fungal Proteins / Q000737:chemistry / Q000378:metabolism*; D008969:Molecular Sequence Data; D016296:Mutagenesis; D010455:Peptides / Q000138:chemical synthesis / Q000737:chemistry / Q000378:metabolism; D010957:Plasmids; D011387:Prokaryotic Cells / Q000378:metabolism; D011485:Protein Binding; D012441:Saccharomyces cerevisiae / Q000235:genetics; D029701:Saccharomyces cerevisiae Proteins; D012697:Serine Endopeptidases; D014157:Transcription Factors / Q000737:chemistry / Q000378:metabolism*; D000099160:LexA Repressor Protein
chemical_list: D000596:Amino Acids; D001426:Bacterial Proteins; D004271:DNA, Fungal; D004268:DNA-Binding Proteins; D005656:Fungal Proteins; D010455:Peptides; D029701:Saccharomyces cerevisiae Proteins; D012697:Serine Endopeptidases; D014157:Transcription Factors; D000099160:LexA Repressor Protein; C052130:GAL4 protein, S cerevisiae