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Attenuation of a DNA Cruciform by a Conserved Regulator Directs T3SS-1 mediated virulence in Vibrio parahaemolyticus

View ORCID ProfileLandon J. Getz, Justin M. Brown, Lauren Sobot, Alexandra Chow, Jastina Mahendrarajah, View ORCID ProfileNikhil A. Thomas
doi: https://doi.org/10.1101/2022.03.07.483294
Landon J. Getz
1Department of Microbiology and Immunology, Faculty of Medicine, Dalhousie University. Halifax, NS
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Justin M. Brown
1Department of Microbiology and Immunology, Faculty of Medicine, Dalhousie University. Halifax, NS
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Lauren Sobot
1Department of Microbiology and Immunology, Faculty of Medicine, Dalhousie University. Halifax, NS
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Alexandra Chow
1Department of Microbiology and Immunology, Faculty of Medicine, Dalhousie University. Halifax, NS
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Jastina Mahendrarajah
1Department of Microbiology and Immunology, Faculty of Medicine, Dalhousie University. Halifax, NS
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Nikhil A. Thomas
1Department of Microbiology and Immunology, Faculty of Medicine, Dalhousie University. Halifax, NS
2Department of Medicine, Faculty of Medicine, Dalhousie University. Halifax, NS
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  • For correspondence: n.thomas@dal.ca
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Abstract

Pathogenic Vibrio species account for 3-5 million annual life-threatening human infections. Virulence is driven by bacterial hemolysin and toxin gene expression often positively regulated by the winged helix-turn-helix (wHTH) HlyU transcriptional regulator family and silenced by Histone-like nucleoid structural protein (H-NS). In the case of Vibrio parahaemolyticus, HlyU is required for virulence gene expression associated with Type 3 Secretion System-1 (T3SS-1) although its mechanism of action is not understood. Here, we provide evidence for DNA cruciform attenuation mediated by HlyU binding to support concomitant virulence gene expression. Genetic and biochemical experiments revealed that upon HlyU mediated DNA cruciform attenuation, an intergenic cryptic promoter became accessible allowing for exsA mRNA expression and initiation of an ExsA autoactivation feedback loop at a separate ExsA-dependent promoter. Using a heterologous E. coli expression system, we reconstituted the dual promoter elements which revealed that HlyU binding and DNA cruciform attenuation were strictly required to initiate the ExsA autoactivation loop. The data indicate that HlyU acts to attenuate a transcriptional repressive DNA cruciform to support T3SS-1 virulence gene expression and reveals a non-canonical extricating gene regulation mechanism in pathogenic Vibrio species.

Competing Interest Statement

The authors have declared no competing interest.

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The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under a CC-BY-NC-ND 4.0 International license.
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Posted March 07, 2022.
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Attenuation of a DNA Cruciform by a Conserved Regulator Directs T3SS-1 mediated virulence in Vibrio parahaemolyticus
Landon J. Getz, Justin M. Brown, Lauren Sobot, Alexandra Chow, Jastina Mahendrarajah, Nikhil A. Thomas
bioRxiv 2022.03.07.483294; doi: https://doi.org/10.1101/2022.03.07.483294
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Attenuation of a DNA Cruciform by a Conserved Regulator Directs T3SS-1 mediated virulence in Vibrio parahaemolyticus
Landon J. Getz, Justin M. Brown, Lauren Sobot, Alexandra Chow, Jastina Mahendrarajah, Nikhil A. Thomas
bioRxiv 2022.03.07.483294; doi: https://doi.org/10.1101/2022.03.07.483294

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