Role of FTL_1306 (dipA) in Francisella tularensis susceptibility to resazomycins.

Authors

  • Kendall Souder West Liberty University
  • Michael Whaby West Liberty University
  • Jennifer Hickman West Liberty University
  • Deanna Scmitt West Liberty University
  • Donald Primerano Marshall University
  • James Denvir Marshall University

Abstract

The CDC classifies Francisella tularensis as a Category A bioterrorism agent. Due to the risk of potential release of antibiotic-resistant F. tularensis strains, new therapeutics against F. tularensis must be developed.  Resazomycins are resazurin (Rz)-based compounds that exhibit antimicrobial activity against F. tularensis and other gram-negative bacteria. The action of resazomycins is not well understood, but potential targets of the antibiotic were identified in a high throughput screen for Rz-resistant isolates. The dipA (FTL_1306) gene was identified as mutated in half of the 48 Rz-resistant (RZR) strains sequenced. To further investigate the role of dipA in Rz susceptibility, we introduced a wild-type copy of dipA into select RZR isolates (RZR1, 5, 43, and 46) that contain dipA mutations. The dipA gene was amplified by PCR from wild-type F. tularensis and cloned into the F. tularensis shuttle vector pABST to generate a construct (pABST-dipA) in which dipA will be constitutively expressed under control of the groEL promoter. The pABST-dipA plasmid was mobilized into each of the selected RZR isolates by electroporation. The resulting RZR dipA-complemented strains were cultivated on chocolate agar containing 10xMIC of Rz to determine their susceptibility to resazomycins. Preliminary results suggest that expression of wild-type dipA in the RZR mutants did not restore sensitivity to Rz indicating dipA may not play a role in Rz susceptibility.  Further investigation is needed to fully elucidate the contribution of dipA to the bactericidal action of resazomycins.

Author Biographies

Kendall Souder, West Liberty University

Department of Natural Sciences and Mathematics

Michael Whaby, West Liberty University

Department of Natural Sciences and Mathematics

Jennifer Hickman, West Liberty University

Department of Natural Sciences and Mathematics

Deanna Scmitt, West Liberty University

Department of Natural Sciences and Mathematics

Donald Primerano, Marshall University

Genomics and Bioinformatics Core Facility

James Denvir, Marshall University

Genomics and Bioinformatics Core Facility

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Published

2020-04-29

How to Cite

Souder, K., Whaby, M., Hickman, J., Scmitt, D., Primerano, D., & Denvir, J. (2020). Role of FTL_1306 (dipA) in Francisella tularensis susceptibility to resazomycins. Proceedings of the West Virginia Academy of Science, 92(1). Retrieved from https://pwvas.org/index.php/pwvas/article/view/654

Issue

Section

Meeting Abstracts-Poster