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Accelerated evolution of a minimal 63-amino acid dual transcription factor

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Broedel, A., Rodrigues, R., Jaramillo, Alfonso and Isalan, M. (2020) Accelerated evolution of a minimal 63-amino acid dual transcription factor. Science Advances, 6 (24). eaba2728. doi:10.1126/sciadv.aba2728

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Official URL: http://dx.doi.org/10.1126/sciadv.aba2728

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Abstract

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 λ is one of the smallest known repressors (66 amino acids), and activators are typically much larger (e.g., λ cI, 237 amino acids). Previous efforts to engineer a minimal activator from λ 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 λ cI 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.

Item Type: Journal Article
Subjects: Q Science > QD Chemistry
Q Science > QP Physiology
Divisions: Faculty of Science > Life Sciences (2010- )
Library of Congress Subject Headings (LCSH): Transcription factors, Bacteriophages -- Genetics, Amino acids, Genetic transcription
Journal or Publication Title: Science Advances
Publisher: American Association for the Advancement of Science
ISSN: 2375-2548
Official Date: 10 June 2020
Dates:
DateEvent
10 June 2020Published
7 April 2020Accepted
Date of first compliant deposit: 11 May 2020
Volume: 6
Number: 24
Article Number: eaba2728
DOI: 10.1126/sciadv.aba2728
Status: Peer Reviewed
Publication Status: Published
Access rights to Published version: Restricted or Subscription Access
RIOXX Funder/Project Grant:
Project/Grant IDRIOXX Funder NameFunder ID
610730, EVOPROGSeventh Framework Programmehttp://dx.doi.org/10.13039/100011102
EVO-ENGINE BB/P020615/1[BBSRC] Biotechnology and Biological Sciences Research Councilhttp://dx.doi.org/10.13039/501100000268
613745, PROMYSFP7 Food, Agriculture and Fisheries, Biotechnologyhttp://dx.doi.org/10.13039/100011262
642738, MetaRNAH2020 Marie Skłodowska-Curie ActionsUNSPECIFIED
BB/M017982/1[EPSRC] Engineering and Physical Sciences Research Councilhttp://dx.doi.org/10.13039/501100000266
BB/M017982/1[BBSRC] Biotechnology and Biological Sciences Research Councilhttp://dx.doi.org/10.13039/501100000268
WT102944Wellcome Trusthttp://dx.doi.org/10.13039/100010269
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