5AKF

THE CRYSTAL STRUCTURE OF I-DMOI Q42AK120M IN COMPLEX WITH ITS TARGET DNA NICKED IN THE CODING STRAND A AND IN THE PRESENCE OF 2MM MN

Summary for 5AKF

• Entry DOI: 10.2210/pdb5akf/pdb
• Related: 5AK9 5AKM 5AKN
• Descriptor: HOMING ENDONUCLEASE I-DMOI, 5'-D(*GP*CP*CP*TP*TP*GP*CP*CP*GP*GP*GP*TP*AP*AP)-3', 5'-D(*GP*TP*TP*CP*CP*GP*GP*CP*GP*CP*GP)-3', ... (8 entities in total)
• Functional Keywords: hydrolase, gene targeting, genetics, protein-dna interaction, homing endonucleases
• Biological source: DESULFUROCOCCUS MOBILIS; More
• Total number of polymer chains: 12
• Total formula weight: 115949.44

Authors

Molina, R.,Marcaida, M.J.,Redondo, P.,Marenchino, M.,D'Abramo, M.,Montoya, G.,Prieto, J. (deposition date: 2015-03-03, release date: 2015-06-17, Last modification date: 2024-01-10)

Primary citation

Molina, R.,Marcaida, M.J.,Redondo, P.,Marenchino, M.,Duchateau, P.,D'Abramo, M.,Montoya, G.,Prieto, J.
Engineering a Nickase on the Homing Endonuclease I-Dmoi Scaffold.
J.Biol.Chem., 290:18534-, 2015

PubMed Abstract: Homing endonucleases are useful tools for genome modification because of their capability to recognize and cleave specifically large DNA targets. These endonucleases generate a DNA double strand break that can be repaired by the DNA damage response machinery. The break can be repaired by homologous recombination, an error-free mechanism, or by non-homologous end joining, a process susceptible to introducing errors in the repaired sequence. The type of DNA cleavage might alter the balance between these two alternatives. The use of "nickases" producing a specific single strand break instead of a double strand break could be an approach to reduce the toxicity associated with non-homologous end joining by promoting the use of homologous recombination to repair the cleavage of a single DNA break. Taking advantage of the sequential DNA cleavage mechanism of I-DmoI LAGLIDADG homing endonuclease, we have developed a new variant that is able to cut preferentially the coding DNA strand, generating a nicked DNA target. Our structural and biochemical analysis shows that by decoupling the action of the catalytic residues acting on each strand we can inhibit one of them while keeping the other functional.

PubMed: 26045557
DOI: 10.1074/JBC.M115.658666

Experimental method

X-RAY DIFFRACTION (2.45 Å)