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Molecular Mechanisms of an All-Purpose Transcription Anti-Termination Factor

47 Pages Posted: 16 Aug 2018 Last revised: 12 Jun 2019 Sneak Peek Status: Published

See all articles by Ferdinand Krupp

Ferdinand Krupp

Charité - Universitätsmedizin Berlin - Department of Medical Physics and Biophysics

Nelly Said

Free University of Berlin (FUB) - Laboratory of Structural Biochemistry

Yong-Heng Huang

Free University of Berlin (FUB) - Laboratory of Structural Biochemistry

Bernhard Loll

Free University of Berlin (FUB) - Laboratory of Structural Biochemistry

Jörg Bürger

Charité - Universitätsmedizin Berlin - Department of Medical Physics and Biophysics

Thorsten Mielke

Free University of Berlin (FUB) - Laboratory of Structural Biochemistry

Christian M. T. Spahn

Charité - Universitätsmedizin Berlin - Department of Medical Physics and Biophysics

Markus Wahl

Free University of Berlin (FUB) - Laboratory of Structural Biochemistry

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Abstract

Bacteriophage λN protein, a paradigmatic anti-termination factor, binds nascent RNA and host Nus factors, rendering RNA polymerase resistant to all pause and termination signals. A 3.7 Å-resolution cryo-electron microscopy structure and structure-informed functional analyses reveal an all-it-takes strategy, in which the intrinsically unstructured λN directly modifies RNA polymerase interactions with the nucleic acids and subverts essential functions of NusA, NusE and NusG to reprogram the transcriptional apparatus. λN repositions NusA and remodels the β-subunit flap tip to preclude folding of pause/termination RNA hairpins in the exit tunnel and disrupt termination-supporting interactions of the α-subunit C-terminal domains. λN invades and traverses the RNA polymerase catalytic cavity, stabilizing the hybrid and impeding pause/termination-related conformational changes. λN also binds upstream DNA, enhancing the weak anti-backtracking and anti-swiveling activities of NusG. Moreover, λN-repositioned NusA and NusE sequester the NusG C-terminal domain, counteracting ρ-dependent termination. Other anti-terminators likely utilize similar mechanisms to enable processive transcription.

Suggested Citation

Krupp, Ferdinand and Said, Nelly and Huang, Yong-Heng and Loll, Bernhard and Bürger, Jörg and Mielke, Thorsten and Spahn, Christian M. T. and Wahl, Markus, Molecular Mechanisms of an All-Purpose Transcription Anti-Termination Factor. Available at SSRN: https://ssrn.com/abstract=3231853 or http://dx.doi.org/10.2139/ssrn.3231853
This is a paper under consideration at Cell Press and has not been peer-reviewed.

Ferdinand Krupp

Charité - Universitätsmedizin Berlin - Department of Medical Physics and Biophysics

Charitéplatz 1
Berlin, 10117
Germany

Nelly Said

Free University of Berlin (FUB) - Laboratory of Structural Biochemistry

Van't-Hoff-Str. 8
Berlin, Berlin 14195
Germany

Yong-Heng Huang

Free University of Berlin (FUB) - Laboratory of Structural Biochemistry

Van't-Hoff-Str. 8
Berlin, Berlin 14195
Germany

Bernhard Loll

Free University of Berlin (FUB) - Laboratory of Structural Biochemistry

Van't-Hoff-Str. 8
Berlin, Berlin 14195
Germany

Jörg Bürger

Charité - Universitätsmedizin Berlin - Department of Medical Physics and Biophysics

Charitéplatz 1
Berlin, 10117
Germany

Thorsten Mielke

Free University of Berlin (FUB) - Laboratory of Structural Biochemistry

Van't-Hoff-Str. 8
Berlin, Berlin 14195
Germany

Christian M. T. Spahn

Charité - Universitätsmedizin Berlin - Department of Medical Physics and Biophysics ( email )

Charitéplatz 1
Berlin, 10117
Germany

Markus Wahl (Contact Author)

Free University of Berlin (FUB) - Laboratory of Structural Biochemistry ( email )

Van't-Hoff-Str. 8
Berlin, Berlin 14195
Germany

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