Non-specific protein-DNA interactions control I-CreI target binding and cleavage

Research output: Contribution to journal › Journal article › Research › peer-review

Standard

Non-specific protein-DNA interactions control I-CreI target binding and cleavage. / Molina, Rafael; Redondo, Pilar; Stella, Stefano; Marenchino, Marco; D'Abramo, Marco; Gervasio, Francesco Luigi; Charles Epinat, Jean; Valton, Julien; Grizot, Silvestre; Duchateau, Phillipe; Prieto, Jesús; Montoya, Guillermo.

In: Nucleic Acids Research, Vol. 40, No. 14, 01.08.2012, p. 6936-6945.

Research output: Contribution to journal › Journal article › Research › peer-review

Harvard

Molina, R, Redondo, P, Stella, S, Marenchino, M, D'Abramo, M, Gervasio, FL, Charles Epinat, J, Valton, J, Grizot, S, Duchateau, P, Prieto, J & Montoya, G 2012, 'Non-specific protein-DNA interactions control I-CreI target binding and cleavage', Nucleic Acids Research, vol. 40, no. 14, pp. 6936-6945. https://doi.org/10.1093/nar/gks320

APA

Molina, R., Redondo, P., Stella, S., Marenchino, M., D'Abramo, M., Gervasio, F. L., Charles Epinat, J., Valton, J., Grizot, S., Duchateau, P., Prieto, J., & Montoya, G. (2012). Non-specific protein-DNA interactions control I-CreI target binding and cleavage. Nucleic Acids Research, 40(14), 6936-6945. https://doi.org/10.1093/nar/gks320

Vancouver

Molina R, Redondo P, Stella S, Marenchino M, D'Abramo M, Gervasio FL et al. Non-specific protein-DNA interactions control I-CreI target binding and cleavage. Nucleic Acids Research. 2012 Aug 1;40(14):6936-6945. https://doi.org/10.1093/nar/gks320

Author

Molina, Rafael ; Redondo, Pilar ; Stella, Stefano ; Marenchino, Marco ; D'Abramo, Marco ; Gervasio, Francesco Luigi ; Charles Epinat, Jean ; Valton, Julien ; Grizot, Silvestre ; Duchateau, Phillipe ; Prieto, Jesús ; Montoya, Guillermo. / Non-specific protein-DNA interactions control I-CreI target binding and cleavage. In: Nucleic Acids Research. 2012 ; Vol. 40, No. 14. pp. 6936-6945.

Bibtex

@article{14608c8123ed47c2a1ea52bc4662c635,
title = "Non-specific protein-DNA interactions control I-CreI target binding and cleavage",
abstract = "Homing endonucleases represent protein scaffolds that provide powerful tools for genome manipulation, as these enzymes possess a very low frequency of DNA cleavage in eukaryotic genomes due to their high specificity. The basis of protein-DNA recognition must be understood to generate tailored enzymes that target the DNA at sites of interest. Protein-DNA interaction engineering of homing endonucleases has demonstrated the potential of these approaches to create new specific instruments to target genes for inactivation or repair. Protein-DNA interface studies have been focused mostly on specific contacts between amino acid side chains and bases to redesign the binding interface. However, it has been shown that 4bp in the central DNA sequence of the 22-bp substrate of a homing endonuclease (I-CreI), which do not show specific protein-DNA interactions, is not devoid of content information. Here, we analyze the mechanism of target discrimination in this substrate region by the I-CreI protein, determining how it can occur independently of the specific protein-DNA interactions. Our data suggest the important role of indirect readout in this substrate region, opening the possibility for a fully rational search of new target sequences, thus improving the development of redesigned enzymes for therapeutic and biotechnological applications.",
author = "Rafael Molina and Pilar Redondo and Stefano Stella and Marco Marenchino and Marco D'Abramo and Gervasio, {Francesco Luigi} and {Charles Epinat}, Jean and Julien Valton and Silvestre Grizot and Phillipe Duchateau and Jes{\'u}s Prieto and Guillermo Montoya",
year = "2012",
month = aug,
day = "1",
doi = "10.1093/nar/gks320",
language = "English",
volume = "40",
pages = "6936--6945",
journal = "Nucleic Acids Research",
issn = "0305-1048",
publisher = "Oxford University Press",
number = "14",

}

RIS

TY - JOUR

T1 - Non-specific protein-DNA interactions control I-CreI target binding and cleavage

AU - Molina, Rafael

AU - Redondo, Pilar

AU - Stella, Stefano

AU - Marenchino, Marco

AU - D'Abramo, Marco

AU - Gervasio, Francesco Luigi

AU - Charles Epinat, Jean

AU - Valton, Julien

AU - Grizot, Silvestre

AU - Duchateau, Phillipe

AU - Prieto, Jesús

AU - Montoya, Guillermo

PY - 2012/8/1

Y1 - 2012/8/1

N2 - Homing endonucleases represent protein scaffolds that provide powerful tools for genome manipulation, as these enzymes possess a very low frequency of DNA cleavage in eukaryotic genomes due to their high specificity. The basis of protein-DNA recognition must be understood to generate tailored enzymes that target the DNA at sites of interest. Protein-DNA interaction engineering of homing endonucleases has demonstrated the potential of these approaches to create new specific instruments to target genes for inactivation or repair. Protein-DNA interface studies have been focused mostly on specific contacts between amino acid side chains and bases to redesign the binding interface. However, it has been shown that 4bp in the central DNA sequence of the 22-bp substrate of a homing endonuclease (I-CreI), which do not show specific protein-DNA interactions, is not devoid of content information. Here, we analyze the mechanism of target discrimination in this substrate region by the I-CreI protein, determining how it can occur independently of the specific protein-DNA interactions. Our data suggest the important role of indirect readout in this substrate region, opening the possibility for a fully rational search of new target sequences, thus improving the development of redesigned enzymes for therapeutic and biotechnological applications.

AB - Homing endonucleases represent protein scaffolds that provide powerful tools for genome manipulation, as these enzymes possess a very low frequency of DNA cleavage in eukaryotic genomes due to their high specificity. The basis of protein-DNA recognition must be understood to generate tailored enzymes that target the DNA at sites of interest. Protein-DNA interaction engineering of homing endonucleases has demonstrated the potential of these approaches to create new specific instruments to target genes for inactivation or repair. Protein-DNA interface studies have been focused mostly on specific contacts between amino acid side chains and bases to redesign the binding interface. However, it has been shown that 4bp in the central DNA sequence of the 22-bp substrate of a homing endonuclease (I-CreI), which do not show specific protein-DNA interactions, is not devoid of content information. Here, we analyze the mechanism of target discrimination in this substrate region by the I-CreI protein, determining how it can occur independently of the specific protein-DNA interactions. Our data suggest the important role of indirect readout in this substrate region, opening the possibility for a fully rational search of new target sequences, thus improving the development of redesigned enzymes for therapeutic and biotechnological applications.

UR - http://www.scopus.com/inward/record.url?scp=84864944748&partnerID=8YFLogxK

U2 - 10.1093/nar/gks320

DO - 10.1093/nar/gks320

M3 - Journal article

C2 - 22495931

AN - SCOPUS:84864944748

VL - 40

SP - 6936

EP - 6945

JO - Nucleic Acids Research

JF - Nucleic Acids Research

SN - 0305-1048

IS - 14

ER -

ID: 202332974