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. 2016 May 12;44(12):5971–5982. doi: 10.1093/nar/gkw423

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© The Author(s) 2016. Published by Oxford University Press on behalf of Nucleic Acids Research.

This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by-nc/4.0/), which permits non-commercial re-use, distribution, and reproduction in any medium, provided the original work is properly cited. For commercial re-use, please contact journals.permissions@oup.com

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Figure 4. — NusG-NTD:NusA-AR2 interaction in the presence of RNAP. (A and B) [¹H,¹⁵N]-HSQC displacement experiment of NusA-AR2 from ¹⁵N-αCTD by NusG-NTD. Black, ¹⁵N-αCTD; red, ¹⁵N-αCTD:NusA-AR2 = 1:1; blue, ¹⁵N-αCTD:NusA-AR2:NusG-NTD = 1:1:3. The concentration of ¹⁵N-αCTD was always 50 μM. The rectangle in (A) indicates the section as in (B). The arrows in (B) show the changes in the chemical shifts of selected residues. (C) NusG binds to NusA in the presence of RNAP. [¹H,¹⁵N]-HSQC spectra of ¹⁵N-NusG, black, ¹⁵N-NusG in the presence of NusA (molar ratio 1:1), red, and ¹⁵N-NusG in the presence of NusA and RNAP (molar ratio 1:1:1), cyan. (D) NusG binds to NusA-AR2 in the presence of RNAP. [¹H,¹⁵N]-HSQC spectra of ¹⁵N-NusG, black, ¹⁵N-NusG in the presence of NusA-AR2 (molar ratio 1:1), red, and ¹⁵N-NusG in the presence of NusA-AR2 and RNAP (molar ratio 1:1:1), cyan. Selected signals are labeled (black, NusG-NTD signals; red, NusG-CTD signals). The concentration of ¹⁵N-NusG was 50 μM in all experiments in (C) and (D). (E) NusA-AR2 removes NusG-NTD from RNAP. [¹H,¹⁵N]-HSQC spectra of ¹⁵N-NusG-NTD, black, ¹⁵N-NusG-NTD in the presence of RNAP (molar ratio 1:0.5), red, and ¹⁵N-NusG-NTD in the presence of RNAP and NusA-AR2 (molar ratio 1:0.5:8), cyan. The concentration of ¹⁵N-NusG was always 50 μM. The rectangle in (E) indicates the section as in (F). In (F), selected signals are assigned with arrows indicating changes in their chemical shifts corresponding to the complex formation of ¹⁵N-NusG-NTD and NusA-AR2.