Work

In-cell architecture of an actively transcribing-translating expressome

Year: 2020

Type: article

Abstract: Structural biology performed inside cells can capture molecular machines in action within their native context. Here we develop an integrative in-cell structural approach using the genome-reduced human pathogen Mycoplasma pneumoniae . We combine whole-cell crosslinking mass spectrometry, cellular cryo-electron tomography, and integrative modeling to determine an in-cell architecture of a transcribing and translating expressome at sub-nanometer resolution. The expressome comprises RNA polymerase (RNAP), the ribosome, and the transcription elongation factors NusG and NusA. We pinpoint NusA at the interface between a NusG-bound elongating RNAP and the ribosome, and propose it could mediate transcription-translation coupling. Translation inhibition dissociates the expressome, whereas transcription inhibition stalls and rearranges it, demonstrating that the active expressome architecture requires both translation and transcription elongation within the cell.

Source: Science

DOI: 10.1126/science.abb3758

Authors: Francis J. O’Reilly, Liang Xue, Andrea Graziadei, Ludwig Sinn, Swantje Lenz, Dimitry Tegunov, Cedric Blötz, Neil Singh, Wim J. H. Hagen, Patrick Cramer, Jörg Stülke, Julia Mahamid, Juri Rappsilber

Institutions: Technische Universität Berlin, Heidelberg University, European Molecular Biology Laboratory, European Molecular Biology Laboratory, Max Planck Institute for Biophysical Chemistry, University of Göttingen, Wellcome Centre for Cell Biology, University of Edinburgh

Language: English

Topic: RNA and protein synthesis mechanisms

Subfield: Molecular Biology

Field: Biochemistry, Genetics and Molecular Biology

Domain: Life Sciences

Open Access status: green

Funders: Wellcome Trust, Deutsche Forschungsgemeinschaft, Bundesministerium für Bildung und Forschung, European Research Council, H2020 European Research Council

Awards: In situ Structural Analysis of Molecular Crowding and Phase Separation, G1117759237, Protein structures in the context of time and space by mass spectrometry., EXC 2008: Unifying Systems in Catalysis "UniSysCat", G1730654401, G2475600967, G2685071433, G3606019796, Erfassung der strukturellen Organisation des Mycoplasma pneumoniae Proteoms mittels inZell CrosslinkingMassenspektrometrie, G4261119002, G5595613502, Structural biology of mammalian transcription regulation, G7987891391, G8136362559, Core Funding for the Wellcome Trust Centre for Cell Biology

OpenAlex ID: W3046695191

FWCI: 12

Cites: 96

Cited by: 267

Related to: 10