Chen, Daiying. Explore the function of KEL-8 in oxidative stress response and search for KEL-8 interacting proteins. Retrieved from https://doi.org/doi:10.7282/T36W9B9H
DescriptionPrevious work in our lab identified KEL-8, a BTB-Kelch superfamily protein, as a substrate receptor for a Cul3 ubiquitin ligase that plays a role in the turnover of GLR-1 AMPA-type glutamate receptors in neurons. C. elegans kel-8 is similar in sequence to Keap1, which encodes a BTB-Kelch protein that negatively regulates the oxidative stress response pathway, leading us to hypothesize that kel-8 is also involved in oxidative stress response, repressing the expression of oxidative stress response genes. We have two major questions: what is the role of KEL-8 in oxidative stress and what are the targets of KEL-8. In order to answer the first question, we used the oxidative stress reporter transgene Pgcs-1::gfp. The gcs-1 gene encodes a phase II detoxification enzyme, γ--glutamine cysteine synthase, and is expressed in the intestine in response to oxidative stress reagents. I tested the expression level of Pgcs-1::gfp in kel-8(od38) nonsense mutants and found that the gcs-1 expression level in kel-8 is constitutively high compared to wild-type animals. Our preliminary results indicate that KEL-8 functions similarly to Keap1 to repress the oxidative stress response. However, the repression phenotype is inconsistent. In addition, we did a yeast two-hybrid screen to search for targets or other interacting proteins of KEL-8, and we found 8 candidates. They are ZC434.8, ric-4, F32D8.12, pccb-1, ssq-4, dim-1, aly-3, and smo-1. Further analyses need to be done to determine whether these are biological meaningful interactions.