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==Cryo-EM structure of E. coli RNA polymerase backtracked elongation complex harboring a terminal mismatch== | |||
<StructureSection load='8fix' size='340' side='right'caption='[[8fix]], [[Resolution|resolution]] 3.90Å' scene=''> | |||
== Structural highlights == | |||
<table><tr><td colspan='2'>[[8fix]] is a 8 chain structure with sequence from [https://en.wikipedia.org/wiki/Escherichia_coli_K-12 Escherichia coli K-12] and [https://en.wikipedia.org/wiki/Escherichia_phage_Lambda Escherichia phage Lambda]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=8FIX OCA]. For a <b>guided tour on the structure components</b> use [https://proteopedia.org/fgij/fg.htm?mol=8FIX FirstGlance]. <br> | |||
</td></tr><tr id='ligand'><td class="sblockLbl"><b>[[Ligand|Ligands:]]</b></td><td class="sblockDat" id="ligandDat"><scene name='pdbligand=MG:MAGNESIUM+ION'>MG</scene>, <scene name='pdbligand=ZN:ZINC+ION'>ZN</scene></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=8fix FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=8fix OCA], [https://pdbe.org/8fix PDBe], [https://www.rcsb.org/pdb/explore.do?structureId=8fix RCSB], [https://www.ebi.ac.uk/pdbsum/8fix PDBsum], [https://prosat.h-its.org/prosat/prosatexe?pdbcode=8fix ProSAT]</span></td></tr> | |||
</table> | |||
<div style="background-color:#fffaf0;"> | |||
== Publication Abstract from PubMed == | |||
In prokaryotes, translation can occur on mRNA that is being transcribed in a process called coupling. How the ribosome affects the RNA polymerase (RNAP) during coupling is not well understood. Here, we reconstituted the E. coli coupling system and demonstrated that the ribosome can prevent pausing and termination of RNAP and double the overall transcription rate at the expense of fidelity. Moreover, we monitored single RNAPs coupled to ribosomes and show that coupling increases the pause-free velocity of the polymerase and that a mechanical assisting force is sufficient to explain the majority of the effects of coupling. Also, by cryo-EM, we observed that RNAPs with a terminal mismatch adopt a backtracked conformation, while a coupled ribosome allosterically induces these polymerases toward a catalytically active anti-swiveled state. Finally, we demonstrate that prolonged RNAP pausing is detrimental to cell viability, which could be prevented by polymerase reactivation through a coupled ribosome. | |||
A trailing ribosome speeds up RNA polymerase at the expense of transcript fidelity via force and allostery.,Wee LM, Tong AB, Florez Ariza AJ, Canari-Chumpitaz C, Grob P, Nogales E, Bustamante CJ Cell. 2023 Mar 16;186(6):1244-1262.e34. doi: 10.1016/j.cell.2023.02.008. PMID:36931247<ref>PMID:36931247</ref> | |||
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.<br> | |||
[[Category: | </div> | ||
[[Category: | <div class="pdbe-citations 8fix" style="background-color:#fffaf0;"></div> | ||
[[Category: | == References == | ||
[[Category: | <references/> | ||
[[Category: Canari | __TOC__ | ||
[[Category: | </StructureSection> | ||
[[Category: | [[Category: Escherichia coli K-12]] | ||
[[Category: | [[Category: Escherichia phage Lambda]] | ||
[[Category: Large Structures]] | |||
[[Category: Bustamante C]] | |||
[[Category: Canari C]] | |||
[[Category: Florez Ariza A]] | |||
[[Category: Grob P]] | |||
[[Category: Nogales E]] | |||
[[Category: Tong A]] | |||
[[Category: Wee L]] | |||
Revision as of 09:52, 29 March 2023
Cryo-EM structure of E. coli RNA polymerase backtracked elongation complex harboring a terminal mismatchCryo-EM structure of E. coli RNA polymerase backtracked elongation complex harboring a terminal mismatch
Structural highlights
Publication Abstract from PubMedIn prokaryotes, translation can occur on mRNA that is being transcribed in a process called coupling. How the ribosome affects the RNA polymerase (RNAP) during coupling is not well understood. Here, we reconstituted the E. coli coupling system and demonstrated that the ribosome can prevent pausing and termination of RNAP and double the overall transcription rate at the expense of fidelity. Moreover, we monitored single RNAPs coupled to ribosomes and show that coupling increases the pause-free velocity of the polymerase and that a mechanical assisting force is sufficient to explain the majority of the effects of coupling. Also, by cryo-EM, we observed that RNAPs with a terminal mismatch adopt a backtracked conformation, while a coupled ribosome allosterically induces these polymerases toward a catalytically active anti-swiveled state. Finally, we demonstrate that prolonged RNAP pausing is detrimental to cell viability, which could be prevented by polymerase reactivation through a coupled ribosome. A trailing ribosome speeds up RNA polymerase at the expense of transcript fidelity via force and allostery.,Wee LM, Tong AB, Florez Ariza AJ, Canari-Chumpitaz C, Grob P, Nogales E, Bustamante CJ Cell. 2023 Mar 16;186(6):1244-1262.e34. doi: 10.1016/j.cell.2023.02.008. PMID:36931247[1] From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine. References
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