DNA-binding miniproteins based on zinc fingers. Assessment of the interaction using nanopores

Jéssica Rodríguez; Soraya Learte-Aymamí; Jesús Mosquera; Garbiñe Celaya; David Rodríguez-Larrea; M. Eugenio Vázquez; José L. Mascareñas

doi:10.1039/c7sc05441f

DNA-binding miniproteins based on zinc fingers. Assessment of the interaction using nanopores

Jéssica Rodríguez, Soraya Learte-Aymamí, Jesús Mosquera, Garbiñe Celaya, David Rodríguez-Larrea, M. Eugenio Vázquez^*, José L. Mascareñas

^*Corresponding author for this work

Pure And Applied Chemistry

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Abstract

Obtaining artificial proteins that mimic the DNA binding properties of natural transcription factors could open new ways of manipulating gene expression at will. In this context it is particularly interesting to develop simple synthetic systems. Inspired by the modularity of natural transcription factors, we have designed synthetic miniproteins that combine the zinc finger module of the transcription factor GAGA and AT-hook peptide domains. These constructs are capable of binding to composite DNA sequences of up to 14 base pairs with high affinity and good selectivity. In particular, we have synthesized three different chimeras and characterized their DNA binding properties by electrophoresis and fluorescence anisotropy. We have also used, for the first time in the study of peptide-based DNA binders, nanopore force spectroscopy to obtain further data on the DNA interaction.

Original language	English
Pages (from-to)	4118-4123
Number of pages	6
Journal	Chemical Science
Volume	9
Issue number	17
Early online date	10 Apr 2018
DOIs	https://doi.org/10.1039/c7sc05441f
Publication status	Published - 7 May 2018

Funding

This work received support from the MINECO (SAF2013-41943-R, BFU2016-81754-ERC, CTQ2015-70698-R, SAF2014-58398-R, and SAF2016-76689-R), the Xunta de Galicia (2015-CP082, ED431C 2017/19 and Centro singular de investigación de Galicia accreditation 2016–2019, ED431G/09), the Fundación AECC (IDEAS197VAZQ), the European Union (European Regional Development Fund – ERDF), and the European Research Council (Advanced Grant No. 340055). Support of the orfeo-cinqa network is also acknowledged. J. R. thanks the Xunta de Galicia for a PhD fellowship; D. R.-L. is a recipient of a Ramón y Cajal Fellowship (RYC-2013-12799).

Keywords

DNA binding
chimeras
DNA interaction
nanopores

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10.1039/c7sc05441fLicence: CC BY 3.0

Rodriguez-etal-CS-2018-DNA-binding-miniproteins-based-on-zinc-fingers-Assessment-of-theFinal published version, 1.43 MBLicence: CC BY 3.0

Cite this

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AU - Celaya, Garbiñe

AU - Rodríguez-Larrea, David

AU - Vázquez, M. Eugenio

AU - Mascareñas, José L.

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DNA-binding miniproteins based on zinc fingers. Assessment of the interaction using nanopores

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