| pubmed-article:21029741 | rdf:type | pubmed:Citation | lld:pubmed |
| pubmed-article:21029741 | lifeskim:mentions | umls-concept:C0205145 | lld:lifeskim |
| pubmed-article:21029741 | lifeskim:mentions | umls-concept:C0524637 | lld:lifeskim |
| pubmed-article:21029741 | lifeskim:mentions | umls-concept:C0012854 | lld:lifeskim |
| pubmed-article:21029741 | lifeskim:mentions | umls-concept:C0014230 | lld:lifeskim |
| pubmed-article:21029741 | lifeskim:mentions | umls-concept:C0205217 | lld:lifeskim |
| pubmed-article:21029741 | lifeskim:mentions | umls-concept:C0015219 | lld:lifeskim |
| pubmed-article:21029741 | lifeskim:mentions | umls-concept:C0037791 | lld:lifeskim |
| pubmed-article:21029741 | pubmed:issue | 1 | lld:pubmed |
| pubmed-article:21029741 | pubmed:dateCreated | 2010-12-21 | lld:pubmed |
| pubmed-article:21029741 | pubmed:abstractText | Elucidating how homing endonucleases undergo changes in recognition site specificity will facilitate efforts to engineer proteins for gene therapy applications. I-SceI is a monomeric homing endonuclease that recognizes and cleaves within an 18-bp target. It tolerates limited degeneracy in its target sequence, including substitution of a C:G(+4) base pair for the wild-type A:T(+4) base pair. Libraries encoding randomized amino acids at I-SceI residue positions that contact or are proximal to A:T(+4) were used in conjunction with a bacterial one-hybrid system to select I-SceI derivatives that bind to recognition sites containing either the A:T(+4) or the C:G(+4) base pairs. As expected, isolates encoding wild-type residues at the randomized positions were selected using either target sequence. All I-SceI proteins isolated using the C:G(+4) recognition site included small side-chain substitutions at G100 and either contained (K86R/G100T, K86R/G100S and K86R/G100C) or lacked (G100A, G100T) a K86R substitution. Interestingly, the binding affinities of the selected variants for the wild-type A:T(+4) target are 4- to 11-fold lower than that of wild-type I-SceI, whereas those for the C:G(+4) target are similar. The increased specificity of the mutant proteins is also evident in binding experiments in vivo. These differences in binding affinities account for the observed ?36-fold difference in target preference between the K86R/G100T and wild-type proteins in DNA cleavage assays. An X-ray crystal structure of the K86R/G100T mutant protein bound to a DNA duplex containing the C:G(+4) substitution suggests how sequence specificity of a homing enzyme can increase. This biochemical and structural analysis defines one pathway by which site specificity is augmented for a homing endonuclease. | lld:pubmed |
| pubmed-article:21029741 | pubmed:grant | http://linkedlifedata.com/r... | lld:pubmed |
| pubmed-article:21029741 | pubmed:language | eng | lld:pubmed |
| pubmed-article:21029741 | pubmed:journal | http://linkedlifedata.com/r... | lld:pubmed |
| pubmed-article:21029741 | pubmed:citationSubset | IM | lld:pubmed |
| pubmed-article:21029741 | pubmed:chemical | http://linkedlifedata.com/r... | lld:pubmed |
| pubmed-article:21029741 | pubmed:chemical | http://linkedlifedata.com/r... | lld:pubmed |
| pubmed-article:21029741 | pubmed:chemical | http://linkedlifedata.com/r... | lld:pubmed |
| pubmed-article:21029741 | pubmed:chemical | http://linkedlifedata.com/r... | lld:pubmed |
| pubmed-article:21029741 | pubmed:chemical | http://linkedlifedata.com/r... | lld:pubmed |
| pubmed-article:21029741 | pubmed:chemical | http://linkedlifedata.com/r... | lld:pubmed |
| pubmed-article:21029741 | pubmed:chemical | http://linkedlifedata.com/r... | lld:pubmed |
| pubmed-article:21029741 | pubmed:status | MEDLINE | lld:pubmed |
| pubmed-article:21029741 | pubmed:month | Jan | lld:pubmed |
| pubmed-article:21029741 | pubmed:issn | 1089-8638 | lld:pubmed |
| pubmed-article:21029741 | pubmed:author | pubmed-author:GimbleFrederi... | lld:pubmed |
| pubmed-article:21029741 | pubmed:author | pubmed-author:HoKwok KiKK | lld:pubmed |
| pubmed-article:21029741 | pubmed:author | pubmed-author:GoldenBarbara... | lld:pubmed |
| pubmed-article:21029741 | pubmed:author | pubmed-author:TenneyKristen... | lld:pubmed |
| pubmed-article:21029741 | pubmed:author | pubmed-author:ChenJui-HuiJH | lld:pubmed |
| pubmed-article:21029741 | pubmed:author | pubmed-author:JoshiRakeshR | lld:pubmed |
| pubmed-article:21029741 | pubmed:copyrightInfo | Copyright © 2010 Elsevier Ltd. All rights reserved. | lld:pubmed |
| pubmed-article:21029741 | pubmed:issnType | Electronic | lld:pubmed |
| pubmed-article:21029741 | pubmed:day | 7 | lld:pubmed |
| pubmed-article:21029741 | pubmed:volume | 405 | lld:pubmed |
| pubmed-article:21029741 | pubmed:owner | NLM | lld:pubmed |
| pubmed-article:21029741 | pubmed:authorsComplete | Y | lld:pubmed |
| pubmed-article:21029741 | pubmed:pagination | 185-200 | lld:pubmed |
| pubmed-article:21029741 | pubmed:dateRevised | 2011-10-6 | lld:pubmed |
| pubmed-article:21029741 | pubmed:meshHeading | pubmed-meshheading:21029741... | lld:pubmed |
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| pubmed-article:21029741 | pubmed:meshHeading | pubmed-meshheading:21029741... | lld:pubmed |
| pubmed-article:21029741 | pubmed:meshHeading | pubmed-meshheading:21029741... | lld:pubmed |
| pubmed-article:21029741 | pubmed:meshHeading | pubmed-meshheading:21029741... | lld:pubmed |
| pubmed-article:21029741 | pubmed:year | 2011 | lld:pubmed |
| pubmed-article:21029741 | pubmed:articleTitle | Evolution of I-SceI homing endonucleases with increased DNA recognition site specificity. | lld:pubmed |
| pubmed-article:21029741 | pubmed:affiliation | Department of Biochemistry, Purdue University, West Lafayette, IN 47907, USA. | lld:pubmed |
| pubmed-article:21029741 | pubmed:publicationType | Journal Article | lld:pubmed |
| pubmed-article:21029741 | pubmed:publicationType | Research Support, U.S. Gov't, Non-P.H.S. | lld:pubmed |
| pubmed-article:21029741 | pubmed:publicationType | Research Support, N.I.H., Extramural | lld:pubmed |
