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Design of a colicin E7 based chimeric zinc-finger nuclease

Colicin E7 is a natural bacterial toxin. Its nuclease domain (NColE7) enters the target cell and kills it by digesting the nucleic acids. The HNH-motif as the catalytic centre of NColE7 at the C-terminus requires the positively charged N-terminal loop for the nuclease activity—offering opportunities...

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Autores principales: Németh, Eszter, Schilli, Gabriella K., Nagy, Gábor, Hasenhindl, Christoph, Gyurcsik, Béla, Oostenbrink, Chris
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Springer International Publishing 2014
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4104000/
https://www.ncbi.nlm.nih.gov/pubmed/24952471
http://dx.doi.org/10.1007/s10822-014-9765-8
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author Németh, Eszter
Schilli, Gabriella K.
Nagy, Gábor
Hasenhindl, Christoph
Gyurcsik, Béla
Oostenbrink, Chris
author_facet Németh, Eszter
Schilli, Gabriella K.
Nagy, Gábor
Hasenhindl, Christoph
Gyurcsik, Béla
Oostenbrink, Chris
author_sort Németh, Eszter
collection PubMed
description Colicin E7 is a natural bacterial toxin. Its nuclease domain (NColE7) enters the target cell and kills it by digesting the nucleic acids. The HNH-motif as the catalytic centre of NColE7 at the C-terminus requires the positively charged N-terminal loop for the nuclease activity—offering opportunities for allosteric control in a NColE7-based artificial nuclease. Accordingly, four novel zinc finger nucleases were designed by computational methods exploiting the special structural features of NColE7. The constructed models were subjected to MD simulations. The comparison of structural stability and functional aspects showed that these models may function as safely controlled artificial nucleases. This study was complemented by random mutagenesis experiments identifying potentially important residues for NColE7 function outside the catalytic region. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1007/s10822-014-9765-8) contains supplementary material, which is available to authorized users.
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spelling pubmed-41040002014-07-21 Design of a colicin E7 based chimeric zinc-finger nuclease Németh, Eszter Schilli, Gabriella K. Nagy, Gábor Hasenhindl, Christoph Gyurcsik, Béla Oostenbrink, Chris J Comput Aided Mol Des Article Colicin E7 is a natural bacterial toxin. Its nuclease domain (NColE7) enters the target cell and kills it by digesting the nucleic acids. The HNH-motif as the catalytic centre of NColE7 at the C-terminus requires the positively charged N-terminal loop for the nuclease activity—offering opportunities for allosteric control in a NColE7-based artificial nuclease. Accordingly, four novel zinc finger nucleases were designed by computational methods exploiting the special structural features of NColE7. The constructed models were subjected to MD simulations. The comparison of structural stability and functional aspects showed that these models may function as safely controlled artificial nucleases. This study was complemented by random mutagenesis experiments identifying potentially important residues for NColE7 function outside the catalytic region. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1007/s10822-014-9765-8) contains supplementary material, which is available to authorized users. Springer International Publishing 2014-06-22 2014 /pmc/articles/PMC4104000/ /pubmed/24952471 http://dx.doi.org/10.1007/s10822-014-9765-8 Text en © The Author(s) 2014 https://creativecommons.org/licenses/by/4.0/ Open AccessThis article is distributed under the terms of the Creative Commons Attribution License which permits any use, distribution, and reproduction in any medium, provided the original author(s) and the source are credited.
spellingShingle Article
Németh, Eszter
Schilli, Gabriella K.
Nagy, Gábor
Hasenhindl, Christoph
Gyurcsik, Béla
Oostenbrink, Chris
Design of a colicin E7 based chimeric zinc-finger nuclease
title Design of a colicin E7 based chimeric zinc-finger nuclease
title_full Design of a colicin E7 based chimeric zinc-finger nuclease
title_fullStr Design of a colicin E7 based chimeric zinc-finger nuclease
title_full_unstemmed Design of a colicin E7 based chimeric zinc-finger nuclease
title_short Design of a colicin E7 based chimeric zinc-finger nuclease
title_sort design of a colicin e7 based chimeric zinc-finger nuclease
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4104000/
https://www.ncbi.nlm.nih.gov/pubmed/24952471
http://dx.doi.org/10.1007/s10822-014-9765-8
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