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 α-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 α-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 regons — these results are consistent with the model that activating regions often form amphipathic α-helices. This work is novel because hydrophobic amino acids are also shown to be important in activating regions of yeast transciption factors.
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References
Brendel V, Karlin S (1989) Association of charge clusters with functional domains of cellular transcription factors. Proc Natl Acad Sci USA 86: 5698–5702
Brent R, Ptashne M (1985) A eukaryotic transcriptional activator bearing the DNA specificity of a prokaryotic repressor. Cell 43: 729–736
Bushman FD, Shang C, Ptashne M (1989) A single glutamic acid residue plays a key role in the transcriptional activation function of lambda repressor. Cell 58: 1163–1171
Carey M, Kakidani H, Leatherwood J, Mostashari F, Ptashne M (1990) An amino-terminal fragment of GAL4 binds DNA as a dimer. J Mol Biol 209: 423–432
Courey AJ, Tjian R (1988) Anaoysis of Sp1 in vivo reveals multiple transcriptional domains, including a novel glutamine-rich activation motif. Cell 55: 887–898
Cress WD, Triezenberg SJ (1991) Critical structural elements of the VP16 transcriptional activation domain. Science 251: 87–90
Driever W, Ma J, Nüsslein-Vohard C, Ptashne M (1989) Rescue of bicoid mutant Drosophila embryos by bicoid fusion proteins containing heterologous activating sequences. Nature 342: 149–154
Frankel AD, Kim PS (1991) Modular structure of transcription factors. Cell 65: 717–719
Gill G, Sadowski I, Ptashne M (1990) Mutations that increase the activity of a transcriptional activator in yeast and mammalian cells. Proc Natl Acad Sci USA 87: 2127–2131
Giniger E, Ptashne M (1987) Transcription in yeast activated by a putative amphipathic alpha helix linked to a DNA binding unit. Nature 330: 670–672
Himmelfarb HJ, Pearlberg J, Last D, Ptashne M (1991) GAL11P: a yeast mutation that potentiates the effect of weak GAL4-derived activators. Cell 63: 1299–1309
Hochschild A, Ptashne M (1988) Interaction at a distance between lambda repressors disrupts gene activation. Nature 336: 353–357
Hope IA, Struhl K (1986) Functional dissection of a eukaryotic transcriptional activator protein, GCN4 of yeast. Cell 46: 885–894
Hope IA, Mahadevan S, Struhl K (1988) Structural and functional characterization of the short acidic transcriptional activation region of yeast GCN4 protein. Nature 333: 635–640
Johnston M (1987) A model fungal gene regulatory circuit: the GAL genes of Saccharomyces cerevisiae. Microbiol Rev 51: 458–476
Lin Y-S, Green MR (1991) Mechanism of action of an acidic transcriptional activator in vitro. Cell 64: 971–981
Little LW, Mount DW (1982) The SOS regulatory system of Escherichia coli. Cell 29: 11–22
Ma J Ptashne M (1987a) Deletion analysis of GAL4 defines two transcriptional activating segments. Cell 48: 847–853
Ma J, Ptashne M (1987b) A new class of yeast transcriptional activators. Cell 51: 113–119
Mermod N, O'Neill EA, Kelly TJ, Tjian R (1988) The proline-rich transcriptional activator of CTF/NF-1 is distinct from the replication and DNA binding domain. Cell 58: 741–753
Mitchell PJ, Tjian R (1989) Transcription regulation in mammalian cells by sequence-specific DNA binding proteins. Science 245: 371–378
Ptashne M (1988) How eukaryotic transcriptional activators work. Nature 335: 683–689
Ptashne M, Gann AF (1990) Activators and targets. Nature 346: 329–331
Ruden DM, Ma J, Ptashne M (1988) No strict alignment is required between a transcriptional activator binding site and the “TATA box” of a yeast gene. Proc Natl Acad Sci USA 85: 4262–4266
Ruden DM, Ma J, Wood K, Li Y, Ptashne M (1991) Generating yeast transcriptional activators containing no yeast protein sequences. Nature 350: 250–252
Sherman F, Fink G, Lawrence C (1983), Methods in yeast genetics, revised. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY
Sigler PB (1988) Acid blobs and negative noodles. Nature 333: 210–213
Stringer KF, Ingles CJ, Greenblatt J (1990) Direct and selective binding of an acidic transcriptional activator domain to the TATA-box factor TFIID. Nature 345: 783–786
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Ruden, D.M. Activating regions of yeast transcription factors must have both acidic and hydrophobic amino acids. Chromosoma 101, 342–348 (1992). https://doi.org/10.1007/BF00346013
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DOI: https://doi.org/10.1007/BF00346013