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

51 Pages Posted: 7 Apr 2020 Publication Status: Published

See all articles by Robert S. Washburn

Robert S. Washburn

Columbia University - Department of Microbiology & Immunology

Philipp K. Zuber

University of Bayreuth

Ming Sun

Columbia University - Department of Biological Sciences

Yaser Hashem

Columbia University - Department of Biochemistry and Molecular Biophysics

Bingxin Shen

Columbia University - Department of Biochemistry and Molecular Biophysics

Wen Li

Columbia University - Department of Biochemistry and Molecular Biophysics

Sho Harvey

California Institute of Technology

Stefan H. Knauer

University of Bayreuth

Joachim Frank

Columbia University - Department of Biological Sciences

Max Gottesman

Columbia University - Department of Microbiology & Immunology

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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.

Keywords: NusG, Transcription: translation coupling, cryo-EM & NMR structures, tmRNA

Suggested Citation

Washburn, Robert S. and Zuber, Philipp K. and Sun, Ming and Hashem, Yaser and Shen, Bingxin and Li, Wen and Harvey, Sho and Knauer, Stefan H. and Frank, Joachim and Gottesman, Max, Escherichia coli NusG Links the Lead Ribosome with the Transcription Elongation Complex. Available at SSRN: https://ssrn.com/abstract=3564986 or http://dx.doi.org/10.2139/ssrn.3564986
This version of the paper has not been formally peer reviewed.

Robert S. Washburn

Columbia University - Department of Microbiology & Immunology

United States

Philipp K. Zuber

University of Bayreuth

Universitatsstr 30
Bayreuth, D-95447
Germany

Ming Sun

Columbia University - Department of Biological Sciences

New York, NY 10027
United States

Yaser Hashem

Columbia University - Department of Biochemistry and Molecular Biophysics

3022 Broadway
New York, NY 10027
United States

Bingxin Shen

Columbia University - Department of Biochemistry and Molecular Biophysics ( email )

3022 Broadway
New York, NY 10027
United States

Wen Li

Columbia University - Department of Biochemistry and Molecular Biophysics

3022 Broadway
New York, NY 10027
United States

Sho Harvey

California Institute of Technology

Pasadena, CA 91125
United States

Stefan H. Knauer

University of Bayreuth

Universitatsstr 30
Bayreuth, D-95447
Germany

Joachim Frank (Contact Author)

Columbia University - Department of Biological Sciences ( email )

New York, NY 10027
United States

Max Gottesman

Columbia University - Department of Microbiology & Immunology

United States

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