2026-06-14T03:31:41Zhttps://repisalud.isciii.es/rest/oai/requestoai:repisalud.isciii.es:20.500.12105/66142024-11-29T17:13:50Zcom_20.500.12105_2173com_20.500.12105_2051col_20.500.12105_19597
Repisalud
author
Prieto Lugo, Francisco Jesus
author
Redondo Bastante, Pilar
author
López-Méndez, Blanca
author
D'Abramo, Marco
author
Merino, Nekane
author
Blanco, Francisco J
author
Duchateau, Phillipe
author
Montoya, Guillermo
author
Molina, Rafael
funder
Ministerio de Ciencia e Innovación (España)
funder
Novo Nordisk Foundation
2018-11-16T10:35:16Z
2018-11-16T10:35:16Z
2018-07-06
Sci Rep. 2018; 8(1):10286.
10.1038/s41598-018-28599-0
2045-2322
2045-2322
Scientific reports
29980759
http://hdl.handle.net/20.500.12105/6614
The high DNA specificity of homing endonucleases makes them a powerful protein scaffold to engineer enzymes for genome manipulation. Understanding their molecular recognition of DNA is an important prerequisite to generate engineered enzymes able to cleave DNA in specific desired genome sites. Protein-DNA recognition studies have been mostly focused on specific direct contacts between amino acid side chains and bases to redesign the binding interface. However, the important role of indirect readout in the central region of the target DNA of the homing endonuclease I-CreI suggested that indirect readout may play a key role in the redesign of protein-DNA interactions. The sequences of the I-CreI central substrate region, 2NN, along with the adjacent 5NNN, are key for substrate cleavage. Here, we analyse the mechanism of target discrimination at the 5NNN region by the I-CreI protein, revealing its critical role in the location and occupancy of the catalytic metal ions, which is crucial for cleavage. Our data highlight the importance of indirect readout for target DNA cleavage, thus aiding I-CreI engineering when targeting new DNA sequences.
eng
HOMING ENDONUCLEASE
SITE RECOGNITION
NUCLEIC-ACIDS
CLEAVAGE
BINDING
SEQUENCES
SOFTWARE
SYSTEM
Understanding the indirect DNA read-out specificity of I-CreI Meganuclease
journal article
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UnderstandingtheindirectDNA_2018.pdf.txt
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