2I3P

K28R mutant of Homing Endonuclease I-CreI

Summary for 2I3P

• Entry DOI: 10.2210/pdb2i3p/pdb
• Related: 2I3Q
• Descriptor: 5'-D(*GP*CP*AP*AP*AP*TP*CP*GP*TP*CP*GP*TP*GP*AP*GP*AP*CP*AP*AP*TP*TP*TP*CP*G)-3', 5'-D(*CP*GP*AP*AP*AP*TP*TP*GP*TP*CP*TP*CP*AP*CP*GP*AP*CP*GP*AP*TP*TP*TP*GP*C)-3', DNA endonuclease I-CreI, ... (5 entities in total)
• Functional Keywords: homing endonulease i-crei, dna, hydrolase-dna complex, hydrolase/dna
• Biological source: Chlamydomonas reinhardtii
• Cellular location: Plastid, chloroplast: P05725
• Total number of polymer chains: 4
• Total formula weight: 50169.36

Authors

Sussman, D.,Rosen, L. (deposition date: 2006-08-20, release date: 2006-09-05, Last modification date: 2023-08-30)

Primary citation

Rosen, L.E.,Morrison, H.A.,Masri, S.,Brown, M.J.,Springstubb, B.,Sussman, D.,Stoddard, B.L.,Seligman, L.M.
Homing endonuclease I-CreI derivatives with novel DNA target specificities.
Nucleic Acids Res., 34:4791-4800, 2006

PubMed Abstract: Homing endonucleases are highly specific enzymes, capable of recognizing and cleaving unique DNA sequences in complex genomes. Since such DNA cleavage events can result in targeted allele-inactivation and/or allele-replacement in vivo, the ability to engineer homing endonucleases matched to specific DNA sequences of interest would enable powerful and precise genome manipulations. We have taken a step-wise genetic approach in analyzing individual homing endonuclease I-CreI protein/DNA contacts, and describe here novel interactions at four distinct target site positions. Crystal structures of two mutant endonucleases reveal the molecular interactions responsible for their altered DNA target specificities. We also combine novel contacts to create an endonuclease with the predicted target specificity. These studies provide important insights into engineering homing endonucleases with novel target specificities, as well as into the evolution of DNA recognition by this fascinating family of proteins.

PubMed: 16971456
DOI: 10.1093/nar/gkl645

Experimental method

X-RAY DIFFRACTION (2.3 Å)