8FIY

Cryo-EM structure of E. coli RNA polymerase Elongation complex in the Transcription-Translation Complex (RNAP in an anti-swiveled conformation)

Summary for 8FIY

• Entry DOI: 10.2210/pdb8fiy/pdb
• EMDB information: 29213
• Descriptor: DNA-directed RNA polymerase subunit alpha, DNA-directed RNA polymerase subunit beta, DNA-directed RNA polymerase subunit beta', ... (8 entities in total)
• Functional Keywords: rna polymerase, ribosome, coupling, transcription, transcription-dna-rna complex, transcription/dna/rna
• Biological source: Escherichia coli K-12; More
• Total number of polymer chains: 7
• Total formula weight: 393971.48

Authors

Florez Ariza, A.,Wee, L.,Tong, A.,Canari, C.,Grob, P.,Nogales, E.,Bustamante, C. (deposition date: 2022-12-17, release date: 2023-03-29, Last modification date: 2025-05-14)

Primary citation

Wee, L.M.,Tong, A.B.,Florez Ariza, A.J.,Canari-Chumpitaz, C.,Grob, P.,Nogales, E.,Bustamante, C.J.
A trailing ribosome speeds up RNA polymerase at the expense of transcript fidelity via force and allostery.
Cell, 186:1244-1262.e34, 2023

PubMed Abstract: 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.

PubMed: 36931247
DOI: 10.1016/j.cell.2023.02.008

Experimental method

ELECTRON MICROSCOPY (7.3 Å)