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Escherichia coli NusG Links the Lead Ribosome with the Transcription Elongation Complex

Washburn, Robert S. and Zuber, Philipp K. and Sun, Ming and Hashem, Yaser and Shen, Bingxin and Li, Wen and Harvey, Sho and Acosta Reyes, Francisco J. and Gottesman, Max E. and Knauer, Stefan H. and Frank, Joachim (2020) Escherichia coli NusG Links the Lead Ribosome with the Transcription Elongation Complex. iScience, 23 (8). Art. No. 101352. ISSN 2589-0042. https://resolver.caltech.edu/CaltechAUTHORS:20191212-105209753

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Abstract

It has been known for more than 50 years that transcription and translation are physically coupled in bacteria, but whether or not this coupling may be mediated by the two-domain protein N-utilization substance (Nus) G in Escherichia coli is still heavily debated. Here, we combine integrative structural biology and functional analyses to provide conclusive evidence that NusG can physically link transcription with translation by contacting both RNA polymerase and the ribosome. We present a cryo-electron microscopy structure of a NusG:70S ribosome complex and nuclear magnetic resonance spectroscopy data revealing simultaneous binding of NusG to RNAP and the intact 70S ribosome, providing the first direct structural evidence for NusG-mediated coupling. Furthermore, in vivo reporter assays show that recruitment of NusG occurs late in transcription and strongly depends on translation. Thus, our data suggest that coupling occurs initially via direct RNAP:ribosome contacts and is then mediated by NusG.


Item Type:Article
Related URLs:
URLURL TypeDescription
https://doi.org/10.1016/j.isci.2020.101352DOIArticle
https://doi.org/10.1101/871962DOIDiscussion Paper
ORCID:
AuthorORCID
Acosta Reyes, Francisco J.0000-0001-8524-1484
Knauer, Stefan H.0000-0002-4143-0694
Additional Information:© 2020 The Authors. This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/). Received 11 March 2020, Revised 4 June 2020, Accepted 3 July 2020, Available online 9 July 2020. We gratefully acknowledge the help of D. Shapoval, M. Bubunenko, N. Costantino, and D. Court, and we are deeply thankful for numerous useful discussions with P. Rösch. Supported by HHMI and NIH R01 GM29169 (to J.F.), NIH R01 GM037219 (to M.E.G.), and the German Research Foundation grant Ro617/21-1 (to P. Rösch). S.H. was supported by an Amgen Fellowship. Author Contributions: B.S., M.S., P.K.Z., R.S.W., S.H., and Y.H. performed the experiments and processed the data. J.F., M.E.G., M.S., P.K.Z., R.S.W., S.H.K., and Y.H. designed the experiments and interpreted the results. F.J.A.R., M.S., and W.L. modeled the atomic structure of 30S:NusG-CTD. J.F., M.E.G., M.S., P.K.Z., and S.H.K. wrote the paper. This publication was funded by the German Research Foundation and the University of Bayreuth in the funding program “Open Access Publishing”. Declaration of Interests: The authors declare no competing financial interests.
Funders:
Funding AgencyGrant Number
Howard Hughes Medical Institute (HHMI)UNSPECIFIED
NIHR01 GM29169
NIHR01 GM037219
Deutsche Forschungsgemeinschaft (DFG)Ro617/21-1
AmgenUNSPECIFIED
University of BayreuthUNSPECIFIED
Subject Keywords:Biochemistry; Biochemistry Methods; Structural Biology; Three-Dimensional Reconstruction of Biomolecular Structures
Issue or Number:8
Record Number:CaltechAUTHORS:20191212-105209753
Persistent URL:https://resolver.caltech.edu/CaltechAUTHORS:20191212-105209753
Official Citation:Robert S. Washburn, Philipp K. Zuber, Ming Sun, Yaser Hashem, Bingxin Shen, Wen Li, Sho Harvey, Francisco J. Acosta Reyes, Max E. Gottesman, Stefan H. Knauer, Joachim Frank, Escherichia coli NusG Links the Lead Ribosome with the Transcription Elongation Complex, iScience, Volume 23, Issue 8, 2020, 101352, ISSN 2589-0042, https://doi.org/10.1016/j.isci.2020.101352. (http://www.sciencedirect.com/science/article/pii/S2589004220305393)
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:100272
Collection:CaltechAUTHORS
Deposited By: George Porter
Deposited On:16 Dec 2019 15:56
Last Modified:08 Oct 2020 21:48

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