2i3p
From Proteopedia
(New page: 200px<br /> <applet load="2i3p" size="450" color="white" frame="true" align="right" spinBox="true" caption="2i3p, resolution 2.300Å" /> '''K28R mutant of Hom...) |
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| - | [[Image:2i3p.gif|left|200px]]<br /> | + | [[Image:2i3p.gif|left|200px]]<br /><applet load="2i3p" size="350" color="white" frame="true" align="right" spinBox="true" |
| - | <applet load="2i3p" size=" | + | |
caption="2i3p, resolution 2.300Å" /> | caption="2i3p, resolution 2.300Å" /> | ||
'''K28R mutant of Homing Endonuclease I-CreI'''<br /> | '''K28R mutant of Homing Endonuclease I-CreI'''<br /> | ||
==Overview== | ==Overview== | ||
| - | Homing endonucleases are highly specific enzymes, capable of recognizing | + | 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. |
==About this Structure== | ==About this Structure== | ||
| - | 2I3P is a [http://en.wikipedia.org/wiki/Single_protein Single protein] structure of sequence from [http://en.wikipedia.org/wiki/Chlamydomonas_reinhardtii Chlamydomonas reinhardtii] with CA as [http://en.wikipedia.org/wiki/ligand ligand]. Full crystallographic information is available from [http:// | + | 2I3P is a [http://en.wikipedia.org/wiki/Single_protein Single protein] structure of sequence from [http://en.wikipedia.org/wiki/Chlamydomonas_reinhardtii Chlamydomonas reinhardtii] with <scene name='pdbligand=CA:'>CA</scene> as [http://en.wikipedia.org/wiki/ligand ligand]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=2I3P OCA]. |
==Reference== | ==Reference== | ||
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[[Category: homing endonulease i-crei]] | [[Category: homing endonulease i-crei]] | ||
| - | ''Page seeded by [http:// | + | ''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Thu Feb 21 17:48:43 2008'' |
Revision as of 15:48, 21 February 2008
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K28R mutant of Homing Endonuclease I-CreI
Overview
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.
About this Structure
2I3P is a Single protein structure of sequence from Chlamydomonas reinhardtii with as ligand. Full crystallographic information is available from OCA.
Reference
Homing endonuclease I-CreI derivatives with novel DNA target specificities., Rosen LE, Morrison HA, Masri S, Brown MJ, Springstubb B, Sussman D, Stoddard BL, Seligman LM, Nucleic Acids Res. 2006;34(17):4791-800. Epub 2006 Sep 13. PMID:16971456
Page seeded by OCA on Thu Feb 21 17:48:43 2008
