[
International Worm Meeting,
2021]
Caenorhabditis elegans is an ideal model organism for studying the xenobiotic detoxification pathways of various natural and synthetic toxins. One such toxin that has been shown to cause death in C. elegans is 1-hydroxyphenazine (1-HP), a molecule produced by the bacterium Pseudomonas aeruginosa. Prior research in our lab has shown the median lethal dose (LD50) for 1-HP in C elegans is 179 muM in PD1074 and between 150-200muM in N2.(Asif et al., 2021; Stupp et al., 2013). Prior research has also shown that C. elegans detoxifies 1-HP by glycosylating it with one, two, or three glucose molecules in N2 worms.(Stupp et al., 2013) We hypothesize that UDP-Glucuronosyltransferase (UGT) enzymes are responsible for glycosylating 1-HP in C. elegans. To identify UGT enzymes implicated in the glycosylation of 1-HP in PD1074, we have implemented our plate-based toxicity assay developed in our prior work on available UGT strains. We began by testing eight UGT mutants, UGT-1, UGT-6, UGT-9, UGT-23, UGT-49, UGT-60, UGT-62, and UGT-66, at the LD50 concentration of 1-HP in PD1074. We screened for mutants with a different mortality rate to N2 and PD1074 worms. Additionally, we will perform HPLC/UV analysis and NMR analysis in order to describe the differences in glycosylation patterns and the ratios of glycosylated and unglycosylated products in mutant strains with differential susceptibility to 1-HP than N2 and PD1074 worms. This could help explain the variation in mortality rates between the different strains and help us understand the complexity of UGTs in C. elegans.
Van der Gaag, Victoria L., Edison, Arthur S., Muzio, Cole J., Asif, Muhammad Zaka, Nocilla, Kelsey A., Guo, Jane
[
MicroPubl Biol,
2021]
1-Hydroxyphenazine (1-HP) is a small molecule produced by Pseudomonas aeruginosa, a bacterium that is used for pathogenesis models in C. elegans (Cezairliyan et al., 2013; Mahajan-Miklos, Tan, Rahme, & Ausubel, 1999). 1-HP is an especially interesting toxin to study as it has been shown to interact with human cells causing ciliary-slowing associated with dyskinesia and ciliostasis (Wilson et al., 1987). Prior research in our lab has shown that this molecule is toxic to C. elegans, with an LD50 between 150 and 200 M, but C. elegans can glycosylate 1-HP, which detoxifies the molecule (Stupp et al., 2013).