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. 2018 Jul 6;8:10286. doi: 10.1038/s41598-018-28599-0

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© The Author(s) 2018

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Active site detailed view of both I-CreI variants in complex with cleavable and non-cleavable targets. (a) Cleavage analysis of I-CreI_D75N_target (left panel) vs I-CreI_D75N_target-null (right panel) and (b) I-CreI_3115_target (left panel) vs I-CreI_3115_target-null (right panel). Omit map (σ = 6.0) around the cleavable phosphodiester revealed no cleavage in the corresponding target-null and phosphodiester cleavage with their original targets. The green spheres represent the Mg²⁺ ions and the red sphere a water molecule. (c) Metal positioning analysis of I-CreI_D75N_target (left panel) vs I-CreI_D75N_target-null (right panel) and (d) I-CreI_3115_target (left panel) vs I-CreI_3115_target-null (right panel). Anomalous maps (σ = 6.0) around the active site revealed the presence of Mn²⁺ (grey spheres) as a catalytic ion with different occupancies depending on the target to cleave. Black arrows indicate the phosphodiester bond cleaved.