Towards a minimal motif for artificial transcriptional activators Journal Article
| Authors: | Ansari, A. Z.; Mapp, A. K.; Nguyen, D. H.; Dervan, P. B.; Ptashne, M. |
| Article Title: | Towards a minimal motif for artificial transcriptional activators |
| Abstract: | Background: Most transcriptional activators minimally comprise two functional modules, one for DNA binding and the other for activation. Several activators also bear an oligomerization region and bind DNA as dimers or higher order oligomers. In a previous study we substituted these domains of a protein activator with synthetic counterparts [Mapp et al., Proc. Natl. Acad. Sci. USA 97 (2000) 3930-3935]. An artificial transcriptional activator, 4.2 kDa in size, comprised of a DNA binding hairpin polyamide tethered to a 20 residue activating peptide (AH) was shown to stimulate promoter specific transcription [Mapp et al., Proc. Natl. Acad. Sci. USA 97 (2000) 3930-3935]. The question arises as to the general nature and the versatility of this minimal activator motif and whether smaller ligands can be designed which maintain potent activation function. Results: Here we have replaced the 20 amino acid AH peptide with eight or 16 residues derived from the activation domain of the potent viral activator VP16. The 16 residue activation module coupled to the polyamide activated transcription over two-fold better than the analogous AH conjugate. Altering the site of attachment of the activation module on the polyamide allowed reduction of the intervening linker from 36 atoms to eight without significant diminution of the activation potential. In this study we also exchanged the polyamide to target a different sequence without compromising the activation function further demonstrating the generality of this design. Conclusions: The polyamide activator conjugates described here represent a class of DNA binding ligands which are tethered to a second functional moiety, viz. an activation domain, that recruits elements of the endogenous transcriptional machinery. Our results define the minimal structural elements required to construct artificial, small molecule activators. If such activators are cell-permeable and can be targeted to designated sites in the genome, this series of conjugates may then serve as a tool to study mechanistic aspects of transcriptional regulation and eventually to modulate gene expression relevant to human diseases. © 2001 Elsevier Science Ltd. |
| Keywords: | gene function; transcription, genetic; gene activation; dna; transcription regulation; amino acid sequence; molecular sequence data; cell membrane; transactivation; base sequence; amino acid; binding sites; trans-activators; dna binding; oligomerization; promoter regions (genetics); polyamide; chemical regulator; hairpin polyamide; small molecule activator; targeted gene control; herpes simplex virus protein vmw65 |
| Journal Title: | Chemistry and Biology |
| Volume: | 8 |
| Issue: | 6 |
| ISSN: | 1074-5521 |
| Publisher: | Elsevier Inc. |
| Date Published: | 2001-06-01 |
| Start Page: | 583 |
| End Page: | 592 |
| Language: | English |
| DOI: | 10.1016/s1074-5521(01)00037-0 |
| PUBMED: | 11410377 |
| PROVIDER: | scopus |
| DOI/URL: | |
| Notes: | Export Date: 21 May 2015 -- Source: Scopus |
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