Asif, Muhammad Zaka et al. (2021) MicroPubl Biol

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).

Asif, Muhammad Zaka et al. (2021) International Worm Meeting "Determining the Effects of 1-Hydroxyphenazine Exposure on UGT Mutants in Caenorhabditis elegans"

Shah, Man, Hafeez, Zaki, Nocilla, Kelsey, Asif, Muhammad Zaka, Edison, Arthur

[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.

Hafeez, Zaki et al. (2021) International Worm Meeting "Investigating the Diversity and Distribution of Caenorhabditis elegans in Georgia"

Shaver, Amanda, Hafeez, Zaki, Edison, Arthur, Anderson, Lindsay, Mohamed, Youssef, Spencer, Deontis, Shah, Man, Asif, Muhammad Zaka, Muzio, Cole

[International Worm Meeting, 2021]

The Caenorhabditis elegans Natural Diversity Resource (CeNDR) maintains a library of strains of C. elegans collected from every continent except Antarctica (Caenorhabditis elegans Natural Diversity Resource [CeNDR], 2020). However, currently, there is only one C. elegans strain from the state of Georgia cataloged in CeNDR (CeNDR, 2020). In an attempt to expand upon the diversity of these collected strains and in an effort to study the distribution of C. elegans in Georgia, we set forth to collect a number of wild nematode isolates. Samples of rotting and decaying vegetation were collected from a variety of locations across Georgia. Data such as temperature, location, and sample type were recorded along with images of each collection site using the Nematode Field Sampling app within the data collection app Fulcrum. Nematodes were isolated from these samples and screened by visual inspection for morphological similarity to C. elegans. Potential C. elegans strains were then cultivated. To confirm that an isolate is indeed C. elegans, we will perform PCR and gel electrophoresis. For worms with an appropriately sized PCR band, we intend to submit samples of the PCR product for Sanger sequencing. We will then use NCBI BLAST to compare the sequencing results of the wild isolates with known species. Finally, we will submit frozen isolates of C. elegans to CeNDR for cataloging and whole-genome sequencing. Caenorhabditis elegans Natural Diversity Resource. (2020, August 30). Global Strain Map [Interactive Map]. Retrieved March 24, 2021 from https://www.elegansvariation.org/strain/global-strain-map

Chism, Kyra et al. (2021) International Worm Meeting "Computational Analysis of UDP-glycosyltransferase Variation across Strains of Caenorhabditis elegans"

Taujale, Rahil, Asif, Muhammad Zaka, Benveniste, Maci, Chism, Kyra, Tucker, Niyelle, Watkins, Rockford, Edison, Arthur, Nicolas, Bailey, Levin, Ari, Johnson, Aleya

[International Worm Meeting, 2021]

Caenorhabditis elegans are simple non-parasitic nematodes with a relatively short life cycle and a wealth of genomic information across multiple databases, making them ideal model organisms. However, little is known about the UDP-glycosyltransferases (UGTs) responsible for their innate detoxification response. UGTs are a large family of phase II enzymes responsible for the glycosylation of small molecules across organisms, thus interacting with small molecules such as toxins in the worms' immediate environment. The Edison Vertically Integrated Projects (VIP) Computational Team is a group of undergraduate students who are working to identify the diversity that exists in UGTs across C. elegans isolates from different geographical locations found in the Caenorhabditis elegans Natural Diversity Resource (CeNDR) database in order to make inferences about their evolutionary relationships and functions. The CeNDR database is a collection of wild isolates of C. elegans and their genomic data found globally used by researchers worldwide. Out of the 250 glycotransferases are responsible for transferring sugar molecules to various substrates, there are about 79 UGTs that transfer sugar molecules to small molecules including toxins. Two approaches were implemented to identify UGTs and make inferences based on their variation. First, we created a catalog of UGTs in the N2 reference strain and used them to create a phylogenetic tree that allowed us to depict the relationships between the UGT protein sequences. For our second approach, we quantified UGT variation using the strains found in the CeNDR database. The results and inferences from this research will help us explore possible functions of UGT genes and improve our understanding of UGT variation in C. elegans.