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==Influenza B polymerase bound to vRNA promoter and capped RNA primer== | |||
<StructureSection load='5msg' size='340' side='right' caption='[[5msg]], [[Resolution|resolution]] 3.80Å' scene=''> | |||
== Structural highlights == | |||
<table><tr><td colspan='2'>[[5msg]] is a 6 chain structure. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=5MSG OCA]. For a <b>guided tour on the structure components</b> use [http://oca.weizmann.ac.il/oca-docs/fgij/fg.htm?mol=5MSG FirstGlance]. <br> | |||
</td></tr><tr id='ligand'><td class="sblockLbl"><b>[[Ligand|Ligands:]]</b></td><td class="sblockDat"><scene name='pdbligand=PO4:PHOSPHATE+ION'>PO4</scene></td></tr> | |||
<tr id='NonStdRes'><td class="sblockLbl"><b>[[Non-Standard_Residue|NonStd Res:]]</b></td><td class="sblockDat"><scene name='pdbligand=M7G:7N-METHYL-8-HYDROGUANOSINE-5-DIPHOSPHATE'>M7G</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='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=5msg FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=5msg OCA], [http://pdbe.org/5msg PDBe], [http://www.rcsb.org/pdb/explore.do?structureId=5msg RCSB], [http://www.ebi.ac.uk/pdbsum/5msg PDBsum], [http://prosat.h-its.org/prosat/prosatexe?pdbcode=5msg ProSAT]</span></td></tr> | |||
</table> | |||
<div style="background-color:#fffaf0;"> | |||
== Publication Abstract from PubMed == | |||
Influenza polymerase replicates, via a complementary RNA intermediate (cRNA), and transcribes the eight viral RNA (vRNA) genome segments. To initiate RNA synthesis it is bound to the conserved 5' and 3' extremities of the vRNA or cRNA (the 'promoter'). 5'-3' base-pairing in the distal promoter region is essential to position the template RNA at the polymerase active site, as shown by a new crystal structure with the 3' end threading through the template entry tunnel. We develop fluorescence polarization assays to quantify initiation of cap-primed (transcription) or unprimed (replication) RNA synthesis by recombinant influenza B polymerase bound to the vRNA or cRNA promoter. The rate-limiting step is formation of a primed initiation complex with minimally ApG required to stabilize the 3' end of the template within the active-site. Polymerase bound to the vRNA promoter initiates RNA synthesis terminally, while the cRNA promoter directs internal initiation at a significantly lower rate. Progression to elongation requires breaking the promoter 5'-3' base-pairing region and favourable compensation by the emerging template-product base-pairs. The RNA synthesis assay is adaptable to high-throughput screening for polymerase inhibitors. In a pilot study, we find that initiation at the cRNA promoter is unusually susceptible to inhibition by 2'F-2'dNTPs. | |||
An in vitro fluorescence based study of initiation of RNA synthesis by influenza B polymerase.,Reich S, Guilligay D, Cusack S Nucleic Acids Res. 2017 Jan 26. pii: gkx043. doi: 10.1093/nar/gkx043. PMID:28126917<ref>PMID:28126917</ref> | |||
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.<br> | |||
[[Category: | </div> | ||
<div class="pdbe-citations 5msg" style="background-color:#fffaf0;"></div> | |||
== References == | |||
<references/> | |||
__TOC__ | |||
</StructureSection> | |||
[[Category: RNA-directed RNA polymerase]] | |||
[[Category: Cusack, S]] | |||
[[Category: Guilligay, D]] | |||
[[Category: Capped rna primer]] | |||
[[Category: Influenza b virus rna-dependent rna polymerase]] | |||
[[Category: Viral protein]] | |||
[[Category: Vrna promoter]] | |||
Revision as of 12:27, 10 March 2017
Influenza B polymerase bound to vRNA promoter and capped RNA primerInfluenza B polymerase bound to vRNA promoter and capped RNA primer
Structural highlights
Publication Abstract from PubMedInfluenza polymerase replicates, via a complementary RNA intermediate (cRNA), and transcribes the eight viral RNA (vRNA) genome segments. To initiate RNA synthesis it is bound to the conserved 5' and 3' extremities of the vRNA or cRNA (the 'promoter'). 5'-3' base-pairing in the distal promoter region is essential to position the template RNA at the polymerase active site, as shown by a new crystal structure with the 3' end threading through the template entry tunnel. We develop fluorescence polarization assays to quantify initiation of cap-primed (transcription) or unprimed (replication) RNA synthesis by recombinant influenza B polymerase bound to the vRNA or cRNA promoter. The rate-limiting step is formation of a primed initiation complex with minimally ApG required to stabilize the 3' end of the template within the active-site. Polymerase bound to the vRNA promoter initiates RNA synthesis terminally, while the cRNA promoter directs internal initiation at a significantly lower rate. Progression to elongation requires breaking the promoter 5'-3' base-pairing region and favourable compensation by the emerging template-product base-pairs. The RNA synthesis assay is adaptable to high-throughput screening for polymerase inhibitors. In a pilot study, we find that initiation at the cRNA promoter is unusually susceptible to inhibition by 2'F-2'dNTPs. An in vitro fluorescence based study of initiation of RNA synthesis by influenza B polymerase.,Reich S, Guilligay D, Cusack S Nucleic Acids Res. 2017 Jan 26. pii: gkx043. doi: 10.1093/nar/gkx043. PMID:28126917[1] From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine. References
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