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Guiding Biomolecular Interactions in Cells Using de Novo Protein - Protein Interfaces

Research output: Contribution to journalArticle

Original languageEnglish
Pages (from-to)1284-1293
Number of pages10
JournalACS Synthetic Biology
Volume8
Issue number6
Early online date6 May 2019
DOIs
DateAccepted/In press - 6 May 2019
DateE-pub ahead of print - 6 May 2019
DatePublished (current) - 21 Jun 2019

Abstract

An improved ability to direct and control biomolecular interactions in living cells would have an impact on synthetic biology. A key issue is the need to introduce interacting components that act orthogonally to endogenous proteomes and interactomes. Here, we show that low-complexity, de novo designed protein–protein interaction (PPI) domains can substitute for natural PPIs and guide engineered protein–DNA interactions in Escherichia coli. Specifically, we use de novo homo- and heterodimeric coiled coils to reconstitute a cytoplasmic split adenylate cyclase, recruit RNA polymerase to a promoter and activate gene expression, and oligomerize both natural and designed DNA-binding domains to repress transcription. Moreover, the stabilities of the heterodimeric coiled coils can be modulated by rational design and, thus, adjust the levels of gene activation and repression in vivo. These experiments demonstrate the possibilities for using designed proteins and interactions to control biomolecular systems such as enzyme cascades and circuits in cells.

    Research areas

  • transcriptional control, TAL effectors, protein−protein interaction, DNA−protein interaction, de novo protein design, α-helical coiled coil, SYNTHETIC BIOLOGY

    Structured keywords

  • BrisSynBio
  • Bristol BioDesign Institute

Documents

Documents

  • Full-text PDF (accepted author manuscript)

    Rights statement: This is the author accepted manuscript (AAM). The final published version (version of record) is available online via American Chemical Society at https://pubs.acs.org/doi/10.1021/acssynbio.8b00501. Please refer to any applicable terms of use of the publisher.

    Accepted author manuscript, 14 MB, PDF document

    Embargo ends: 6/05/20

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DOI

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