2xwh: Difference between revisions
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==HCV-J6 NS5B POLYMERASE STRUCTURE AT 1.8 ANGSTROM== | ==HCV-J6 NS5B POLYMERASE STRUCTURE AT 1.8 ANGSTROM== | ||
<StructureSection load='2xwh' size='340' side='right' caption='[[2xwh]], [[Resolution|resolution]] 1.80Å' scene=''> | <StructureSection load='2xwh' size='340' side='right' caption='[[2xwh]], [[Resolution|resolution]] 1.80Å' scene=''> | ||
== Structural highlights == | == Structural highlights == | ||
<table><tr><td colspan='2'>[[2xwh]] is a 1 chain structure with sequence from [http://en.wikipedia.org/wiki/ | <table><tr><td colspan='2'>[[2xwh]] is a 1 chain structure with sequence from [http://en.wikipedia.org/wiki/Hcv Hcv]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=2XWH OCA]. For a <b>guided tour on the structure components</b> use [http://oca.weizmann.ac.il/oca-docs/fgij/fg.htm?mol=2XWH FirstGlance]. <br> | ||
</td></tr><tr id='ligand'><td class="sblockLbl"><b>[[Ligand|Ligands:]]</b></td><td class="sblockDat"><scene name='pdbligand=15P:POLYETHYLENE+GLYCOL+(N=34)'>15P</scene>, <scene name='pdbligand=PEG:DI(HYDROXYETHYL)ETHER'>PEG</scene></td></tr> | </td></tr><tr id='ligand'><td class="sblockLbl"><b>[[Ligand|Ligands:]]</b></td><td class="sblockDat"><scene name='pdbligand=15P:POLYETHYLENE+GLYCOL+(N=34)'>15P</scene>, <scene name='pdbligand=PEG:DI(HYDROXYETHYL)ETHER'>PEG</scene></td></tr> | ||
<tr id='activity'><td class="sblockLbl"><b>Activity:</b></td><td class="sblockDat"><span class='plainlinks'>[http://en.wikipedia.org/wiki/RNA-directed_RNA_polymerase RNA-directed RNA polymerase], with EC number [http://www.brenda-enzymes.info/php/result_flat.php4?ecno=2.7.7.48 2.7.7.48] </span></td></tr> | <tr id='activity'><td class="sblockLbl"><b>Activity:</b></td><td class="sblockDat"><span class='plainlinks'>[http://en.wikipedia.org/wiki/RNA-directed_RNA_polymerase RNA-directed RNA polymerase], with EC number [http://www.brenda-enzymes.info/php/result_flat.php4?ecno=2.7.7.48 2.7.7.48] </span></td></tr> | ||
<tr id='resources'><td class="sblockLbl"><b>Resources:</b></td><td class="sblockDat"><span class='plainlinks'>[http://oca.weizmann.ac.il/oca-docs/fgij/fg.htm?mol=2xwh FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=2xwh OCA], [http://www.rcsb.org/pdb/explore.do?structureId=2xwh RCSB], [http://www.ebi.ac.uk/pdbsum/2xwh PDBsum]</span></td></tr> | <tr id='resources'><td class="sblockLbl"><b>Resources:</b></td><td class="sblockDat"><span class='plainlinks'>[http://oca.weizmann.ac.il/oca-docs/fgij/fg.htm?mol=2xwh FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=2xwh OCA], [http://pdbe.org/2xwh PDBe], [http://www.rcsb.org/pdb/explore.do?structureId=2xwh RCSB], [http://www.ebi.ac.uk/pdbsum/2xwh PDBsum], [http://prosat.h-its.org/prosat/prosatexe?pdbcode=2xwh ProSAT]</span></td></tr> | ||
</table> | </table> | ||
<div style="background-color:#fffaf0;"> | <div style="background-color:#fffaf0;"> | ||
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From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.<br> | From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.<br> | ||
</div> | </div> | ||
<div class="pdbe-citations 2xwh" style="background-color:#fffaf0;"></div> | |||
==See Also== | ==See Also== | ||
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__TOC__ | __TOC__ | ||
</StructureSection> | </StructureSection> | ||
[[Category: Hcv]] | |||
[[Category: RNA-directed RNA polymerase]] | [[Category: RNA-directed RNA polymerase]] | ||
[[Category: Bressanelli, S]] | [[Category: Bressanelli, S]] | ||
[[Category: Scrima, N]] | [[Category: Scrima, N]] | ||
[[Category: Replication]] | [[Category: Replication]] | ||
[[Category: Transferase]] | [[Category: Transferase]] | ||
Revision as of 23:41, 11 August 2016
HCV-J6 NS5B POLYMERASE STRUCTURE AT 1.8 ANGSTROMHCV-J6 NS5B POLYMERASE STRUCTURE AT 1.8 ANGSTROM
Structural highlights
Publication Abstract from PubMedThe HCV genotype 2a isolate JFH1 represents the only cloned HCV wild-type sequence capable of efficient replication in cell culture as well as in vivo. Previous reports have pointed to NS5B, the viral RNA dependent RNA polymerase (RdRp), as a major determinant for efficient replication of this isolate. To understand the contribution of the JFH1 NS5B gene at the molecular level, we aimed at conferring JFH1 properties to NS5B from the closely related J6 isolate. We created intragenotypic chimeras in the NS5B region of JFH1 and J6 and compared replication efficiency in cell culture and RdRp activity of the purified proteins in vitro, revealing more than three independent mechanisms conferring the role of JFH1 NS5B in efficient RNA replication. Most critical was residue I405 in the thumb domain of the polymerase, strongly stimulating replication in cell culture by enhancing overall de novo RNA synthesis. A structural comparison of JFH1 and J6 at high resolution indicated a clear correlation of a closed thumb conformation of the RdRp and the efficiency of the enzyme at de novo RNA synthesis, in accordance with the proposal that I405 enhances de novo initiation. In addition, we identified several residues enhancing replication independent of RdRp activity in vitro. The functional properties of JFH1 NS5B could be restored by a few single nucleotide substitutions to the J6 isolate. Finally, we were able to enhance replication efficiency of a genotype 1b isolate with the I405 mutation, indicating that this mechanism of action is conserved across genotypes. A comprehensive structure-function comparison of hepatitis C virus strains JFH1 and J6 polymerases reveals a key residue stimulating replication in cell culture across genotypes.,Schmitt M, Scrima N, Radujkovic D, Caillet-Saguy C, Simister PC, Friebe P, Wicht O, Klein R, Bartenschlager R, Lohmann V, Bressanelli S J Virol. 2011 Jan 5. PMID:21209117[1] From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine. See AlsoReferences
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