1u0c
From Proteopedia
(New page: 200px<br /> <applet load="1u0c" size="450" color="white" frame="true" align="right" spinBox="true" caption="1u0c, resolution 2.5Å" /> '''Y33C Mutatant of Hom...) |
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| - | [[Image:1u0c.gif|left|200px]]<br /> | + | [[Image:1u0c.gif|left|200px]]<br /><applet load="1u0c" size="350" color="white" frame="true" align="right" spinBox="true" |
| - | <applet load="1u0c" size=" | + | |
caption="1u0c, resolution 2.5Å" /> | caption="1u0c, resolution 2.5Å" /> | ||
'''Y33C Mutatant of Homing endonuclease I-CreI'''<br /> | '''Y33C Mutatant of Homing endonuclease I-CreI'''<br /> | ||
==Overview== | ==Overview== | ||
| - | Homing endonucleases are highly specific DNA endonucleases, encoded within | + | Homing endonucleases are highly specific DNA endonucleases, encoded within mobile introns or inteins, that induce targeted recombination, double-strand repair and gene conversion of their cognate target sites. Due to their biological function and high level of target specificity, these enzymes are under intense investigation as tools for gene targeting. These studies require that naturally occurring enzymes be redesigned to recognize novel target sites. Here, we report studies in which the homodimeric LAGLIDADG homing endonuclease I-CreI is altered at individual side-chains corresponding to contact points to distinct base-pairs in its target site. The resulting enzyme constructs drive specific elimination of selected DNA targets in vivo and display shifted specificities of DNA binding and cleavage in vitro. Crystal structures of two of these constructs demonstrate that substitution of individual side-chain/DNA contact patterns can occur with almost no structural deformation or rearrangement of the surrounding complex, facilitating an isolated, modular redesign strategy for homing endonuclease activity and specificity. |
==About this Structure== | ==About this Structure== | ||
| - | 1U0C 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 MG as [http://en.wikipedia.org/wiki/ligand ligand]. Full crystallographic information is available from [http:// | + | 1U0C 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=MG:'>MG</scene> as [http://en.wikipedia.org/wiki/ligand ligand]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=1U0C OCA]. |
==Reference== | ==Reference== | ||
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[[Category: Fauce, S.]] | [[Category: Fauce, S.]] | ||
[[Category: Monnat, R.]] | [[Category: Monnat, R.]] | ||
| - | [[Category: Seligman, L | + | [[Category: Seligman, L M.]] |
| - | [[Category: Stoddard, B | + | [[Category: Stoddard, B L.]] |
[[Category: Sussman, D.]] | [[Category: Sussman, D.]] | ||
[[Category: MG]] | [[Category: MG]] | ||
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[[Category: protein/dna]] | [[Category: protein/dna]] | ||
| - | ''Page seeded by [http:// | + | ''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Thu Feb 21 15:19:15 2008'' |
Revision as of 13:19, 21 February 2008
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Y33C Mutatant of Homing endonuclease I-CreI
Overview
Homing endonucleases are highly specific DNA endonucleases, encoded within mobile introns or inteins, that induce targeted recombination, double-strand repair and gene conversion of their cognate target sites. Due to their biological function and high level of target specificity, these enzymes are under intense investigation as tools for gene targeting. These studies require that naturally occurring enzymes be redesigned to recognize novel target sites. Here, we report studies in which the homodimeric LAGLIDADG homing endonuclease I-CreI is altered at individual side-chains corresponding to contact points to distinct base-pairs in its target site. The resulting enzyme constructs drive specific elimination of selected DNA targets in vivo and display shifted specificities of DNA binding and cleavage in vitro. Crystal structures of two of these constructs demonstrate that substitution of individual side-chain/DNA contact patterns can occur with almost no structural deformation or rearrangement of the surrounding complex, facilitating an isolated, modular redesign strategy for homing endonuclease activity and specificity.
About this Structure
1U0C is a Single protein structure of sequence from Chlamydomonas reinhardtii with as ligand. Full crystallographic information is available from OCA.
Reference
Isolation and characterization of new homing endonuclease specificities at individual target site positions., Sussman D, Chadsey M, Fauce S, Engel A, Bruett A, Monnat R Jr, Stoddard BL, Seligman LM, J Mol Biol. 2004 Sep 3;342(1):31-41. PMID:15313605
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