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Expression of Bacillus subtilis ABCF antibiotic resistance factor VmlR is regulated by RNA polymerase pausing, transcription attenuation, translation attenuation and (p)ppGpp

Hiraku, Takada LU ; Mandell, Zachary F ; Yakhnin, Helen ; Glazyrina, Anastasiya ; Chiba, Shinobu ; Kurata, Tatsuaki LU ; Wu, Kelvin J Y ; Tresco, Ben I C ; Myers, Andrew G and Atkinson, Gemma LU , et al. (2022) In Nucleic Acids Research 50(11). p.6174-6189
Abstract

Since antibiotic resistance is often associated with a fitness cost, bacteria employ multi-layered regulatory mechanisms to ensure that expression of resistance factors is restricted to times of antibiotic challenge. In Bacillus subtilis, the chromosomally-encoded ABCF ATPase VmlR confers resistance to pleuromutilin, lincosamide and type A streptogramin translation inhibitors. Here we show that vmlR expression is regulated by translation attenuation and transcription attenuation mechanisms. Antibiotic-induced ribosome stalling during translation of an upstream open reading frame in the vmlR leader region prevents formation of an anti-antiterminator structure, leading to the formation of an antiterminator structure that prevents... (More)

Since antibiotic resistance is often associated with a fitness cost, bacteria employ multi-layered regulatory mechanisms to ensure that expression of resistance factors is restricted to times of antibiotic challenge. In Bacillus subtilis, the chromosomally-encoded ABCF ATPase VmlR confers resistance to pleuromutilin, lincosamide and type A streptogramin translation inhibitors. Here we show that vmlR expression is regulated by translation attenuation and transcription attenuation mechanisms. Antibiotic-induced ribosome stalling during translation of an upstream open reading frame in the vmlR leader region prevents formation of an anti-antiterminator structure, leading to the formation of an antiterminator structure that prevents intrinsic termination. Thus, transcription in the presence of antibiotic induces vmlR expression. We also show that NusG-dependent RNA polymerase pausing in the vmlR leader prevents leaky expression in the absence of antibiotic. Furthermore, we demonstrate that induction of VmlR expression by compromised protein synthesis does not require the ability of VmlR to rescue the translational defect, as exemplified by constitutive induction of VmlR by ribosome assembly defects. Rather, the specificity of induction is determined by the antibiotic's ability to stall the ribosome on the regulatory open reading frame located within the vmlR leader. Finally, we demonstrate the involvement of (p)ppGpp-mediated signalling in antibiotic-induced VmlR expression.

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organization
publishing date
type
Contribution to journal
publication status
published
subject
in
Nucleic Acids Research
volume
50
issue
11
article number
gkac497
pages
6174 - 6189
publisher
Oxford University Press
external identifiers
  • scopus:85136156064
  • pmid:35699226
ISSN
1362-4962
DOI
10.1093/nar/gkac497
language
English
LU publication?
yes
id
4fcb1eac-5bf0-42bb-9252-ad5134e08b8b
date added to LUP
2022-06-20 09:19:57
date last changed
2024-04-18 06:46:39
@article{4fcb1eac-5bf0-42bb-9252-ad5134e08b8b,
  abstract     = {{<p>Since antibiotic resistance is often associated with a fitness cost, bacteria employ multi-layered regulatory mechanisms to ensure that expression of resistance factors is restricted to times of antibiotic challenge. In Bacillus subtilis, the chromosomally-encoded ABCF ATPase VmlR confers resistance to pleuromutilin, lincosamide and type A streptogramin translation inhibitors. Here we show that vmlR expression is regulated by translation attenuation and transcription attenuation mechanisms. Antibiotic-induced ribosome stalling during translation of an upstream open reading frame in the vmlR leader region prevents formation of an anti-antiterminator structure, leading to the formation of an antiterminator structure that prevents intrinsic termination. Thus, transcription in the presence of antibiotic induces vmlR expression. We also show that NusG-dependent RNA polymerase pausing in the vmlR leader prevents leaky expression in the absence of antibiotic. Furthermore, we demonstrate that induction of VmlR expression by compromised protein synthesis does not require the ability of VmlR to rescue the translational defect, as exemplified by constitutive induction of VmlR by ribosome assembly defects. Rather, the specificity of induction is determined by the antibiotic's ability to stall the ribosome on the regulatory open reading frame located within the vmlR leader. Finally, we demonstrate the involvement of (p)ppGpp-mediated signalling in antibiotic-induced VmlR expression.</p>}},
  author       = {{Hiraku, Takada and Mandell, Zachary F and Yakhnin, Helen and Glazyrina, Anastasiya and Chiba, Shinobu and Kurata, Tatsuaki and Wu, Kelvin J Y and Tresco, Ben I C and Myers, Andrew G and Atkinson, Gemma and Babitzke, Paul and Hauryliuk, Vasili}},
  issn         = {{1362-4962}},
  language     = {{eng}},
  month        = {{06}},
  number       = {{11}},
  pages        = {{6174--6189}},
  publisher    = {{Oxford University Press}},
  series       = {{Nucleic Acids Research}},
  title        = {{Expression of Bacillus subtilis ABCF antibiotic resistance factor VmlR is regulated by RNA polymerase pausing, transcription attenuation, translation attenuation and (p)ppGpp}},
  url          = {{http://dx.doi.org/10.1093/nar/gkac497}},
  doi          = {{10.1093/nar/gkac497}},
  volume       = {{50}},
  year         = {{2022}},
}