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New page: left|200px<br /><applet load="2ccz" size="450" color="white" frame="true" align="right" spinBox="true" caption="2ccz, resolution 2.70Å" /> '''CRYSTAL STRUCTURE OF...
 
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[[Image:2ccz.gif|left|200px]]<br /><applet load="2ccz" size="450" color="white" frame="true" align="right" spinBox="true"
caption="2ccz, resolution 2.70&Aring;" />
'''CRYSTAL STRUCTURE OF E. COLI PRIMOSOMOL PROTEIN PRIB BOUND TO SSDNA'''<br />


==Overview==
==Crystal structure of E. coli primosomol protein PriB bound to ssDNA==
PriB is a primosomal protein required for replication restart in, Escherichia coli. PriB stimulates PriA helicase activity via interaction, with single-stranded DNA (ssDNA), but the molecular details of this, interaction remain unclear. Here, we report the crystal structure of PriB, complexed with a 15 bases oligonucleotide (dT15) at 2.7 A resolution. PriB, shares structural similarity with the E.coli ssDNA-binding protein, (EcoSSB). However, the structure of the PriB-dT15 complex reveals that, PriB binds ssDNA differently. Results from filter-binding assays show that, PriB-ssDNA interaction is salt-sensitive and cooperative. Mutational, analysis suggests that the loop L45 plays an important role in ssDNA, binding. Based on the crystal structure and biochemical analyses, we, propose a cooperative mechanism for the binding of PriB to ssDNA and a, model for the assembly of the PriA-PriB-ssDNA complex. This report, presents the first structure of a replication restart primosomal protein, complexed with DNA, and a novel model that explains the interactions, between a dimeric oligonucleotide-binding-fold protein and ssDNA.
<StructureSection load='2ccz' size='340' side='right'caption='[[2ccz]], [[Resolution|resolution]] 2.70&Aring;' scene=''>
== Structural highlights ==
<table><tr><td colspan='2'>[[2ccz]] is a 3 chain structure with sequence from [https://en.wikipedia.org/wiki/Escherichia_coli_K-12 Escherichia coli K-12]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=2CCZ OCA]. For a <b>guided tour on the structure components</b> use [https://proteopedia.org/fgij/fg.htm?mol=2CCZ FirstGlance]. <br>
</td></tr><tr id='method'><td class="sblockLbl"><b>[[Empirical_models|Method:]]</b></td><td class="sblockDat" id="methodDat">X-ray diffraction, [[Resolution|Resolution]] 2.7&#8491;</td></tr>
<tr id='resources'><td class="sblockLbl"><b>Resources:</b></td><td class="sblockDat"><span class='plainlinks'>[https://proteopedia.org/fgij/fg.htm?mol=2ccz FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=2ccz OCA], [https://pdbe.org/2ccz PDBe], [https://www.rcsb.org/pdb/explore.do?structureId=2ccz RCSB], [https://www.ebi.ac.uk/pdbsum/2ccz PDBsum], [https://prosat.h-its.org/prosat/prosatexe?pdbcode=2ccz ProSAT]</span></td></tr>
</table>
== Function ==
[https://www.uniprot.org/uniprot/PRIB_ECOLI PRIB_ECOLI] Binds single-stranded DNA at the primosome assembly site (PAS). During primosome assembly it facilitates the complex formation between PriA and DnaT.<ref>PMID:1856227</ref> <ref>PMID:1646811</ref> <ref>PMID:8366072</ref>
== Evolutionary Conservation ==
[[Image:Consurf_key_small.gif|200px|right]]
Check<jmol>
  <jmolCheckbox>
    <scriptWhenChecked>; select protein; define ~consurf_to_do selected; consurf_initial_scene = true; script "/wiki/ConSurf/cc/2ccz_consurf.spt"</scriptWhenChecked>
    <scriptWhenUnchecked>script /wiki/extensions/Proteopedia/spt/initialview01.spt</scriptWhenUnchecked>
    <text>to colour the structure by Evolutionary Conservation</text>
  </jmolCheckbox>
</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=2ccz ConSurf].
<div style="clear:both"></div>
<div style="background-color:#fffaf0;">
== Publication Abstract from PubMed ==
PriB is a primosomal protein required for replication restart in Escherichia coli. PriB stimulates PriA helicase activity via interaction with single-stranded DNA (ssDNA), but the molecular details of this interaction remain unclear. Here, we report the crystal structure of PriB complexed with a 15 bases oligonucleotide (dT15) at 2.7 A resolution. PriB shares structural similarity with the E.coli ssDNA-binding protein (EcoSSB). However, the structure of the PriB-dT15 complex reveals that PriB binds ssDNA differently. Results from filter-binding assays show that PriB-ssDNA interaction is salt-sensitive and cooperative. Mutational analysis suggests that the loop L45 plays an important role in ssDNA binding. Based on the crystal structure and biochemical analyses, we propose a cooperative mechanism for the binding of PriB to ssDNA and a model for the assembly of the PriA-PriB-ssDNA complex. This report presents the first structure of a replication restart primosomal protein complexed with DNA, and a novel model that explains the interactions between a dimeric oligonucleotide-binding-fold protein and ssDNA.


==About this Structure==
Complexed crystal structure of replication restart primosome protein PriB reveals a novel single-stranded DNA-binding mode.,Huang CY, Hsu CH, Sun YJ, Wu HN, Hsiao CD Nucleic Acids Res. 2006;34(14):3878-86. Epub 2006 Aug 9. PMID:16899446<ref>PMID:16899446</ref>
2CCZ is a [http://en.wikipedia.org/wiki/Single_protein Single protein] structure of sequence from [http://en.wikipedia.org/wiki/Escherichia_coli Escherichia coli]. Full crystallographic information is available from [http://ispc.weizmann.ac.il/oca-bin/ocashort?id=2CCZ OCA].


==Reference==
From MEDLINE&reg;/PubMed&reg;, a database of the U.S. National Library of Medicine.<br>
Complexed crystal structure of replication restart primosome protein PriB reveals a novel single-stranded DNA-binding mode., Huang CY, Hsu CH, Sun YJ, Wu HN, Hsiao CD, Nucleic Acids Res. 2006;34(14):3878-86. Epub 2006 Aug 9. PMID:[http://ispc.weizmann.ac.il//pmbin/getpm?pmid=16899446 16899446]
</div>
[[Category: Escherichia coli]]
<div class="pdbe-citations 2ccz" style="background-color:#fffaf0;"></div>
[[Category: Single protein]]
== References ==
[[Category: Hsiao, C.D.]]
<references/>
[[Category: Hsu, C.H.]]
__TOC__
[[Category: Huang, C.Y.]]
</StructureSection>
[[Category: Sun, Y.J.]]
[[Category: Escherichia coli K-12]]
[[Category: Wu, H.N.]]
[[Category: Large Structures]]
[[Category: dna recombination]]
[[Category: Hsiao C-D]]
[[Category: dna repair]]
[[Category: Hsu C-H]]
[[Category: dna replication]]
[[Category: Huang C-Y]]
[[Category: dna/protein complex]]
[[Category: Sun Y-J]]
[[Category: prib]]
[[Category: Wu H-N]]
[[Category: primosome]]
[[Category: single-stranded dna]]
[[Category: ssdna]]
 
''Page seeded by [http://ispc.weizmann.ac.il/oca OCA ] on Wed Nov 21 09:05:19 2007''

Latest revision as of 14:13, 13 December 2023

Crystal structure of E. coli primosomol protein PriB bound to ssDNA

2ccz, resolution 2.70Å

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