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rdf:type |
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lifeskim:mentions |
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pubmed:issue |
3
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pubmed:dateCreated |
2003-8-13
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pubmed:abstractText |
Cre recombinase exchanges DNA strands at the LoxP recognition site via transphosphorylation reactions that involve pentacoordinate transition states. We demonstrate that meta-vanadate ion (VO(3)(-)) and appropriate DNA substrates assemble a transition-state analog-like complex in the Cre active site. Meta-vanadate inhibits recombination of LoxP-derived oligonucleotide substrates that contain a gap at either or both scissile phosphates, but does not inhibit reactions with intact LoxP. The 3(')-hydroxyl group of the gapped substrate is required for inhibition, suggesting that vanadate is ligated by three oxo ligands. Assembly of the inhibited complex is slow (t(1/2)=19min at 4mM NaVO(3)) and requires Cre, substrates, and meta-vanadate. Holliday junction intermediates accumulated at lower meta-vanadate concentrations, suggesting that the second strand exchange is inhibited more readily than the first. The apparent K(D) for meta-vanadate is 1.5-2mM and binding shows positive cooperativity. This methodology may have general application for mechanistic studies of recombinase/topoisomerase-mediated strand exchange reactions.
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pubmed:grant |
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pubmed:commentsCorrections |
http://linkedlifedata.com/resource/pubmed/commentcorrection/12914783-10085061,
http://linkedlifedata.com/resource/pubmed/commentcorrection/12914783-10686599,
http://linkedlifedata.com/resource/pubmed/commentcorrection/12914783-10911997,
http://linkedlifedata.com/resource/pubmed/commentcorrection/12914783-11340053,
http://linkedlifedata.com/resource/pubmed/commentcorrection/12914783-11601846,
http://linkedlifedata.com/resource/pubmed/commentcorrection/12914783-11756402,
http://linkedlifedata.com/resource/pubmed/commentcorrection/12914783-12051940,
http://linkedlifedata.com/resource/pubmed/commentcorrection/12914783-12376595,
http://linkedlifedata.com/resource/pubmed/commentcorrection/12914783-12470949,
http://linkedlifedata.com/resource/pubmed/commentcorrection/12914783-12618186,
http://linkedlifedata.com/resource/pubmed/commentcorrection/12914783-12779336,
http://linkedlifedata.com/resource/pubmed/commentcorrection/12914783-1943774,
http://linkedlifedata.com/resource/pubmed/commentcorrection/12914783-2514790,
http://linkedlifedata.com/resource/pubmed/commentcorrection/12914783-2821547,
http://linkedlifedata.com/resource/pubmed/commentcorrection/12914783-3280015,
http://linkedlifedata.com/resource/pubmed/commentcorrection/12914783-3856690,
http://linkedlifedata.com/resource/pubmed/commentcorrection/12914783-502865,
http://linkedlifedata.com/resource/pubmed/commentcorrection/12914783-5432063,
http://linkedlifedata.com/resource/pubmed/commentcorrection/12914783-6460113,
http://linkedlifedata.com/resource/pubmed/commentcorrection/12914783-6574501,
http://linkedlifedata.com/resource/pubmed/commentcorrection/12914783-6954485,
http://linkedlifedata.com/resource/pubmed/commentcorrection/12914783-8168503,
http://linkedlifedata.com/resource/pubmed/commentcorrection/12914783-8564811,
http://linkedlifedata.com/resource/pubmed/commentcorrection/12914783-8611530,
http://linkedlifedata.com/resource/pubmed/commentcorrection/12914783-8637902,
http://linkedlifedata.com/resource/pubmed/commentcorrection/12914783-9038180,
http://linkedlifedata.com/resource/pubmed/commentcorrection/12914783-9529259,
http://linkedlifedata.com/resource/pubmed/commentcorrection/12914783-9568707,
http://linkedlifedata.com/resource/pubmed/commentcorrection/12914783-9759501
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pubmed:language |
eng
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pubmed:journal |
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pubmed:citationSubset |
IM
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pubmed:chemical |
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pubmed:status |
MEDLINE
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pubmed:month |
Aug
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pubmed:issn |
0006-291X
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pubmed:author |
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pubmed:issnType |
Print
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pubmed:day |
29
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pubmed:volume |
308
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pubmed:owner |
NLM
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pubmed:authorsComplete |
Y
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pubmed:pagination |
529-34
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pubmed:dateRevised |
2010-12-3
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pubmed:meshHeading |
pubmed-meshheading:12914783-Base Sequence,
pubmed-meshheading:12914783-Binding Sites,
pubmed-meshheading:12914783-Dose-Response Relationship, Drug,
pubmed-meshheading:12914783-Enzyme Inhibitors,
pubmed-meshheading:12914783-Integrases,
pubmed-meshheading:12914783-Kinetics,
pubmed-meshheading:12914783-Models, Chemical,
pubmed-meshheading:12914783-Molecular Sequence Data,
pubmed-meshheading:12914783-Recombination, Genetic,
pubmed-meshheading:12914783-Vanadates,
pubmed-meshheading:12914783-Viral Proteins
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pubmed:year |
2003
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pubmed:articleTitle |
Vanadate-based transition-state analog inhibitors of Cre-LoxP recombination.
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pubmed:affiliation |
Section of Molecular and Cellular Biology, University of California, Davis 95616, USA.
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pubmed:publicationType |
Journal Article,
Research Support, Non-U.S. Gov't
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