Figure - PMC

Skip to main content

An official website of the United States government

Here's how you know

Here's how you know

Official websites use .gov
A .gov website belongs to an official government organization in the United States.

Secure .gov websites use HTTPS
A lock ( ) or https:// means you've safely connected to the .gov website. Share sensitive information only on official, secure websites.

View full-text article in PMC

. 2022 Jun 14;50(11):6174–6189. doi: 10.1093/nar/gkac497

Search in PMC
Search in PubMed
View in NLM Catalog
Add to search

© The Author(s) 2022. 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 (https://creativecommons.org/licenses/by/4.0/), which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited.

PMC Copyright notice

Figure 4. — NusG controls lincomycin-dependent induction of VmlR expression. (A, B) Term-seq and RNA-seq (3,47) as well as RNET-seq (3) in wild-type (PLBS730 grown with IPTG), nusA_dep (PLBS730 grown without IPTG), and ΔnusG (PLBS731 grown with IPTG) strains as indicated; ectopic expression of nusA is controlled by an IPTG-inducible P_hy-spankpromoter. (A) The intrinsic terminator in the vmlR leader region requires both NusA and NusG for efficient termination. The NusG-dependent and NusG-independent pauses are marked. The arrow at the top indicates the direction of transcription. (B) Zoomed in magnification showing the NusG-dependent and NusG-independent pauses. (C, D) In vitro reconstitution of NusG-dependent RNAP pausing and NusA/NusG-stimulated termination that controls VmlR expression. In vitro transcription termination assays using DNA templates with wild-type (TTATTT, C) or mutant (TTACAA, (D) NusG pause motifs were performed in the absence or presence of NusA, NusG, or both. Mutations in red are in the consensus NusG-dependent pause motif. Percent termination is shown below each of the 30 min chase reaction lanes. (E) VmlR expression is induced by sub-MIC concentrations of lincomycin and the basal expression of VmlR is repressed by NusG. NusG-deficient (ΔnusG) strain encoding His₆-tagged VmlR (VHB489) was treated with increasing concentrations of lincomycin for 30 min (T30) and VmlR_His6 was detected by α-His₆ immunoblotting. (F) NusG recognizes the TTNTTT motif in the vmlR leader region to repress VmlR expression. Wild-type (VHB223), ΔnusG (VHB489), SL-2* (VHB612) and TTNTTT* (VHB611) strains were treated with 0.4 μg/ml lincomycin for 30 min (T30) and levels of VmlR_His6 were assessed by α-His₆ immunoblotting. Mutations of SL-2* and TTNTTT* are highlighted in red.