2hoi

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[[Image:2hoi.gif|left|200px]]<br /><applet load="2hoi" size="350" color="white" frame="true" align="right" spinBox="true"
 
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caption="2hoi, resolution 2.601&Aring;" />
 
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'''Crystal structure of the tetrameric pre-cleavage synaptic complex in the cre-loxp site-specific recombination'''<br />
 
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==Overview==
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==Crystal structure of the tetrameric pre-cleavage synaptic complex in the cre-loxp site-specific recombination==
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Cre recombinase catalyzes site-specific recombination between 34-bp loxP, sites in a variety of topological and cellular contexts. An obligatory, step in the recombination reaction is the association, or synapsis, of, Cre-bound loxP sites to form a tetrameric protein assembly that is, competent for strand exchange. Using analytical ultracentrifugation and, electrophoresis approaches, we have studied the energetics of Cre-mediated, synapsis of loxP sites. We found that synapsis occurs with a high affinity, (Kd = 10 nM) and is pH-dependent but does not require divalent cations., Surprisingly, the catalytically inactive Cre K201A mutant is fully, competent for synapsis of loxP sites, yet the inactive Y324F and R173K, mutants are defective for synapsis. These findings have allowed us to, determine the first crystal structures of a pre-cleavage Cre-loxP synaptic, complex in a configuration representing the starting point in the, recombination pathway. When combined with a quantitative analysis of, synapsis using loxP mutants, the structures explain how the central 8 bp, of the loxP site are able to dictate the order of strand exchange in the, Cre system.
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<StructureSection load='2hoi' size='340' side='right'caption='[[2hoi]], [[Resolution|resolution]] 2.60&Aring;' scene=''>
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== Structural highlights ==
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<table><tr><td colspan='2'>[[2hoi]] is a 8 chain structure with sequence from [https://en.wikipedia.org/wiki/Escherichia_virus_P1 Escherichia virus P1]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=2HOI OCA]. For a <b>guided tour on the structure components</b> use [https://proteopedia.org/fgij/fg.htm?mol=2HOI FirstGlance]. <br>
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</td></tr><tr id='method'><td class="sblockLbl"><b>[[Empirical_models|Method:]]</b></td><td class="sblockDat" id="methodDat">X-ray diffraction, [[Resolution|Resolution]] 2.601&#8491;</td></tr>
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<tr id='resources'><td class="sblockLbl"><b>Resources:</b></td><td class="sblockDat"><span class='plainlinks'>[https://proteopedia.org/fgij/fg.htm?mol=2hoi FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=2hoi OCA], [https://pdbe.org/2hoi PDBe], [https://www.rcsb.org/pdb/explore.do?structureId=2hoi RCSB], [https://www.ebi.ac.uk/pdbsum/2hoi PDBsum], [https://prosat.h-its.org/prosat/prosatexe?pdbcode=2hoi ProSAT]</span></td></tr>
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</table>
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== Function ==
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[https://www.uniprot.org/uniprot/RECR_BPP1 RECR_BPP1] Catalyzes site-specific recombination between two 34-base-pair LOXP sites. Its role is to maintain the phage genome as a monomeric unit-copy plasmid in the lysogenic state.
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== Evolutionary Conservation ==
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[[Image:Consurf_key_small.gif|200px|right]]
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Check<jmol>
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<jmolCheckbox>
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<scriptWhenChecked>; select protein; define ~consurf_to_do selected; consurf_initial_scene = true; script "/wiki/ConSurf/ho/2hoi_consurf.spt"</scriptWhenChecked>
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<scriptWhenUnchecked>script /wiki/extensions/Proteopedia/spt/initialview01.spt</scriptWhenUnchecked>
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<text>to colour the structure by Evolutionary Conservation</text>
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</jmolCheckbox>
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</jmol>, as determined by [http://consurfdb.tau.ac.il/ ConSurfDB]. You may read the [[Conservation%2C_Evolutionary|explanation]] of the method and the full data available from [http://bental.tau.ac.il/new_ConSurfDB/main_output.php?pdb_ID=2hoi ConSurf].
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<div style="clear:both"></div>
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<div style="background-color:#fffaf0;">
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== Publication Abstract from PubMed ==
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Cre recombinase catalyzes site-specific recombination between 34-bp loxP sites in a variety of topological and cellular contexts. An obligatory step in the recombination reaction is the association, or synapsis, of Cre-bound loxP sites to form a tetrameric protein assembly that is competent for strand exchange. Using analytical ultracentrifugation and electrophoresis approaches, we have studied the energetics of Cre-mediated synapsis of loxP sites. We found that synapsis occurs with a high affinity (Kd = 10 nM) and is pH-dependent but does not require divalent cations. Surprisingly, the catalytically inactive Cre K201A mutant is fully competent for synapsis of loxP sites, yet the inactive Y324F and R173K mutants are defective for synapsis. These findings have allowed us to determine the first crystal structures of a pre-cleavage Cre-loxP synaptic complex in a configuration representing the starting point in the recombination pathway. When combined with a quantitative analysis of synapsis using loxP mutants, the structures explain how the central 8 bp of the loxP site are able to dictate the order of strand exchange in the Cre system.
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==About this Structure==
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Synapsis of loxP sites by Cre recombinase.,Ghosh K, Guo F, Van Duyne GD J Biol Chem. 2007 Aug 17;282(33):24004-16. Epub 2007 Jun 15. PMID:17573343<ref>PMID:17573343</ref>
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2HOI is a [http://en.wikipedia.org/wiki/Single_protein Single protein] structure of sequence from [http://en.wikipedia.org/wiki/Leuconostoc_phage_p37 Leuconostoc phage p37]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=2HOI OCA].
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==Reference==
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From MEDLINE&reg;/PubMed&reg;, a database of the U.S. National Library of Medicine.<br>
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Synapsis of loxP sites by Cre recombinase., Ghosh K, Guo F, Van Duyne GD, J Biol Chem. 2007 Aug 17;282(33):24004-16. Epub 2007 Jun 15. PMID:[http://ispc.weizmann.ac.il//pmbin/getpm?pmid=17573343 17573343]
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</div>
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[[Category: Leuconostoc phage p37]]
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<div class="pdbe-citations 2hoi" style="background-color:#fffaf0;"></div>
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[[Category: Single protein]]
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[[Category: Duyne, G.D.Van.]]
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[[Category: Ghosh, K.]]
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[[Category: cre-loxp precleavage synapse]]
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[[Category: recombination/dna complex]]
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''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Wed Jan 23 12:31:46 2008''
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==See Also==
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*[[Resolvase 3D structures|Resolvase 3D structures]]
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== References ==
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<references/>
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__TOC__
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</StructureSection>
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[[Category: Escherichia virus P1]]
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[[Category: Large Structures]]
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[[Category: Ghosh K]]
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[[Category: Van Duyne GD]]

Current revision

Crystal structure of the tetrameric pre-cleavage synaptic complex in the cre-loxp site-specific recombination

PDB ID 2hoi

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