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''' | ==Crystal structure of the homing endonuclease I-CvuI in complex with its target (Sro1.3) in the presence of 2 mM Ca== | ||
<StructureSection load='5a72' size='340' side='right' caption='[[5a72]], [[Resolution|resolution]] 2.60Å' scene=''> | |||
== Structural highlights == | |||
<table><tr><td colspan='2'>[[5a72]] is a 4 chain structure. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=5A72 OCA]. For a <b>guided tour on the structure components</b> use [http://oca.weizmann.ac.il/oca-docs/fgij/fg.htm?mol=5A72 FirstGlance]. <br> | |||
</td></tr><tr id='ligand'><td class="sblockLbl"><b>[[Ligand|Ligands:]]</b></td><td class="sblockDat"><scene name='pdbligand=CA:CALCIUM+ION'>CA</scene></td></tr> | |||
<tr id='related'><td class="sblockLbl"><b>[[Related_structure|Related:]]</b></td><td class="sblockDat">[[5a73|5a73]], [[5a74|5a74]], [[5a75|5a75]], [[5a77|5a77]], [[5a78|5a78]]</td></tr> | |||
<tr id='resources'><td class="sblockLbl"><b>Resources:</b></td><td class="sblockDat"><span class='plainlinks'>[http://oca.weizmann.ac.il/oca-docs/fgij/fg.htm?mol=5a72 FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=5a72 OCA], [http://pdbe.org/5a72 PDBe], [http://www.rcsb.org/pdb/explore.do?structureId=5a72 RCSB], [http://www.ebi.ac.uk/pdbsum/5a72 PDBsum]</span></td></tr> | |||
</table> | |||
== Function == | |||
[[http://www.uniprot.org/uniprot/DNE1_CHLVU DNE1_CHLVU]] Probable endonuclease involved in intron homing. | |||
<div style="background-color:#fffaf0;"> | |||
== Publication Abstract from PubMed == | |||
Homing endonucleases recognize and generate a DNA double-strand break, which has been used to promote gene targeting. These enzymes recognize long DNA stretches; they are highly sequence-specific enzymes and display a very low frequency of cleavage even in complete genomes. Although a large number of homing endonucleases have been identified, the landscape of possible target sequences is still very limited to cover the complexity of the whole eukaryotic genome. Therefore, the finding and molecular analysis of homing endonucleases identified but not yet characterized may widen the landscape of possible target sequences. The previous characterization of protein-DNA interaction before the engineering of new homing endonucleases is essential for further enzyme modification. Here we report the crystal structure of I-CvuI in complex with its target DNA and with the target DNA of I-CreI, a homologue enzyme widely used in genome engineering. To characterize the enzyme cleavage mechanism, we have solved the I-CvuI DNA structures in the presence of non-catalytic (Ca(2+)) and catalytic ions (Mg(2+)). We have also analyzed the metal dependence of DNA cleavage using Mg(2+) ions at different concentrations ranging from non-cleavable to cleavable concentrations obtained from in vitro cleavage experiments. The structure of I-CvuI homing endonuclease expands the current repertoire for engineering custom specificities, both by itself as a new scaffold alone and in hybrid constructs with other related homing endonucleases or other DNA-binding protein templates. | |||
Crystal Structure of the Homing Endonuclease I-CvuI Provides a New Template for Genome Modification.,Molina R, Redondo P, Lopez-Mendez B, Villate M, Merino N, Blanco FJ, Valton J, Grizot S, Duchateau P, Prieto J, Montoya G J Biol Chem. 2015 Nov 27;290(48):28727-36. doi: 10.1074/jbc.M115.678342. Epub, 2015 Sep 11. PMID:26363068<ref>PMID:26363068</ref> | |||
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.<br> | |||
</div> | |||
<div class="pdbe-citations 5a72" style="background-color:#fffaf0;"></div> | |||
[[Category: | == References == | ||
<references/> | |||
__TOC__ | |||
</StructureSection> | |||
[[Category: Blanco, F J]] | |||
[[Category: Duchateau, P]] | |||
[[Category: Grizot, S]] | |||
[[Category: LopezMendez, B]] | |||
[[Category: Merino, N]] | |||
[[Category: Molina, R]] | [[Category: Molina, R]] | ||
[[Category: | [[Category: Montoya, G]] | ||
[[Category: Prieto, J]] | [[Category: Prieto, J]] | ||
[[Category: Redondo, P]] | [[Category: Redondo, P]] | ||
[[Category: Valton, J]] | [[Category: Valton, J]] | ||
[[Category: Villate, M]] | |||
[[Category: Gene targeting]] | |||
[[Category: Genetic]] | |||
[[Category: Homing endonuclease]] | |||
[[Category: Hydrolase-dna complex]] | |||
[[Category: Protein-dna interaction]] | |||
Revision as of 10:32, 9 December 2015
Crystal structure of the homing endonuclease I-CvuI in complex with its target (Sro1.3) in the presence of 2 mM CaCrystal structure of the homing endonuclease I-CvuI in complex with its target (Sro1.3) in the presence of 2 mM Ca
Structural highlights
Function[DNE1_CHLVU] Probable endonuclease involved in intron homing. Publication Abstract from PubMedHoming endonucleases recognize and generate a DNA double-strand break, which has been used to promote gene targeting. These enzymes recognize long DNA stretches; they are highly sequence-specific enzymes and display a very low frequency of cleavage even in complete genomes. Although a large number of homing endonucleases have been identified, the landscape of possible target sequences is still very limited to cover the complexity of the whole eukaryotic genome. Therefore, the finding and molecular analysis of homing endonucleases identified but not yet characterized may widen the landscape of possible target sequences. The previous characterization of protein-DNA interaction before the engineering of new homing endonucleases is essential for further enzyme modification. Here we report the crystal structure of I-CvuI in complex with its target DNA and with the target DNA of I-CreI, a homologue enzyme widely used in genome engineering. To characterize the enzyme cleavage mechanism, we have solved the I-CvuI DNA structures in the presence of non-catalytic (Ca(2+)) and catalytic ions (Mg(2+)). We have also analyzed the metal dependence of DNA cleavage using Mg(2+) ions at different concentrations ranging from non-cleavable to cleavable concentrations obtained from in vitro cleavage experiments. The structure of I-CvuI homing endonuclease expands the current repertoire for engineering custom specificities, both by itself as a new scaffold alone and in hybrid constructs with other related homing endonucleases or other DNA-binding protein templates. Crystal Structure of the Homing Endonuclease I-CvuI Provides a New Template for Genome Modification.,Molina R, Redondo P, Lopez-Mendez B, Villate M, Merino N, Blanco FJ, Valton J, Grizot S, Duchateau P, Prieto J, Montoya G J Biol Chem. 2015 Nov 27;290(48):28727-36. doi: 10.1074/jbc.M115.678342. Epub, 2015 Sep 11. PMID:26363068[1] From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine. References
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