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University of Washington

1. Tabakh, Hannah. Characterizing the mechanisms of resistance to the novel antimicrobial 4-hydroxy-2-nonenal by the model pathogen Listeria monocytogenes.

Degree: PhD, 2021, University of Washington

Pathogens encounter numerous antimicrobial responses, including the reactive oxygen species (ROS) burst. ROS-mediated oxidation of host membrane poly-unsaturated fatty acids (PUFAs) generates the toxic ab-carbonyl 4-hydroxy-2-nonenal (4-HNE). Though studied extensively in the context of sterile inflammation, 4-HNE’s role during infection remains limited. Here we found that 4-HNE is generated during bacterial infection and that the intracellular pathogen Listeria monocytogenes induces a specific set of genes in response to 4-HNE exposure. A component of the L. monocytogenes 4-HNE response is the expression of the genes rha1 and rha2 which code for two NADPH-dependent oxidoreductases that collectively counter 4-HNE toxicity by converting 4-HNE to the less toxic product 4-hydroxynonanal (4-HNA). Heterologous expression of rha1/2 in Bacillus subtilis significantly increased bacterial resistance to 4-HNE both in vitro and following phagocytosis by murine macrophages. Our work demonstrates that 4-HNE is a previously unappreciated component of ROS-mediated toxicity and that L. monocytogenes has evolved specific countermeasures to survive within its presence. In addition to the above-mentioned work, we also developed a saturated transposon library in L. monocytogenes that we then used to determine the essential genes of L. monocytogenes using a transposon sequencing (Tn-seq) approach. This methodology could be used in the future to further probe the genetic determinants of L. monocytogenes resistance to 4-HNE. Advisors/Committee Members: Woodward, Joshua J (advisor).

Subjects/Keywords: 4-HNE; 4-hydroxy-2-nonenal; 4-hydroxynonenal; Listeria monocytogenes; reactive oxygen species; ROS; Microbiology; Biology; Microbiology

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APA · Chicago · MLA · Vancouver · CSE | Export to Zotero / EndNote / Reference Manager

APA (6th Edition):

Tabakh, H. (2021). Characterizing the mechanisms of resistance to the novel antimicrobial 4-hydroxy-2-nonenal by the model pathogen Listeria monocytogenes. (Doctoral Dissertation). University of Washington. Retrieved from http://hdl.handle.net/1773/46849

Chicago Manual of Style (16th Edition):

Tabakh, Hannah. “Characterizing the mechanisms of resistance to the novel antimicrobial 4-hydroxy-2-nonenal by the model pathogen Listeria monocytogenes.” 2021. Doctoral Dissertation, University of Washington. Accessed April 22, 2021. http://hdl.handle.net/1773/46849.

MLA Handbook (7th Edition):

Tabakh, Hannah. “Characterizing the mechanisms of resistance to the novel antimicrobial 4-hydroxy-2-nonenal by the model pathogen Listeria monocytogenes.” 2021. Web. 22 Apr 2021.

Vancouver:

Tabakh H. Characterizing the mechanisms of resistance to the novel antimicrobial 4-hydroxy-2-nonenal by the model pathogen Listeria monocytogenes. [Internet] [Doctoral dissertation]. University of Washington; 2021. [cited 2021 Apr 22]. Available from: http://hdl.handle.net/1773/46849.

Council of Science Editors:

Tabakh H. Characterizing the mechanisms of resistance to the novel antimicrobial 4-hydroxy-2-nonenal by the model pathogen Listeria monocytogenes. [Doctoral Dissertation]. University of Washington; 2021. Available from: http://hdl.handle.net/1773/46849

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