Journal
SCIENCE ADVANCES
Volume 6, Issue 24, Pages -Publisher
AMER ASSOC ADVANCEMENT SCIENCE
DOI: 10.1126/sciadv.aba2728
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Funding
- European Commission [610730]
- BBSRC grant EVO-ENGINE [BB/P020615/1]
- FP7-KBBE [613745]
- EPSRC-BBSRC [BB/M017982/1]
- Wellcome Trust U.K [WT102944]
- Volkswagen Foundation
- [642738]
- BBSRC [BB/P020615/1] Funding Source: UKRI
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Transcription factors control gene expression in all life. This raises the question of what is the smallest protein that can support such activity. In nature, Cro from bacteriophage lambda is one of the smallest known repressors (66 amino acids), and activators are typically much larger (e.g., lambda cl, 237 amino acids). Previous efforts to engineer a minimal activator from lambda Cro resulted in no activity in vivo in cells. In this study, we show that directed evolution results in a new Cro activator-repressor that functions as efficiently as lambda cl in vivo. To achieve this, we develop phagemid-assisted continuous evolution (PACEmid). We find that a peptide as small as 63 amino acids functions efficiently as an activator and/or repressor. To our knowledge, this is the smallest protein activator that enables polymerase recruitment, highlighting the capacity of transcription factors to evolve from very short peptide sequences.
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