Genetic analysis of Caenorhabditis elegans pry-1/Axin suppressors identifies genes involved in reproductive structure development, stress responses, and aging

Abstract

The Axin family of scaffolding proteins regulates a wide array of developmental and post-developmental processes in eukaryotes. Studies in the nematode Caenorhabditis elegans have shown that the Axin homolog PRY-1 plays essential roles in multiple tissues. To understand the genetic network of pry-1, we focused on a set of genes that are differentially expressed in the pry-1-mutant transcriptome and are linked to reproductive structure development. Knocking down eight of the genes (spp-1, clsp-1, ard-1, rpn-7, cpz-1, his-7, cdk-1, and rnr-1) via RNA interference efficiently suppressed the multivulva phenotype of pry-1 mutants. In all cases, the ectopic induction of P3.p vulval precursor cell was also inhibited. The suppressor genes are members of known gene families in eukaryotes and perform essential functions. Our genetic interaction experiments revealed that in addition to their role in vulval development, these genes participate in one or more pry-1-mediated biological events. Whereas four of them (cpz-1, his-7, cdk-1, and rnr-1) function in both stress response and aging, two (spp-1 and ard-1) are specific to stress response. Altogether, these findings demonstrate the important role of pry-1 suppressors in regulating developmental and post-developmental processes in C. elegans. Given that the genes described in this study are conserved, future investigations of their interactions with Axin and their functional specificity promises to uncover the genetic network of Axin in metazoans.

The FGFR4 Homolog KIN-9 Regulates Lifespan and Stress Responses in Caenorhabditis elegans

“…PRY-1 also regulates the CABIN1 domain-containing protein, PICD-1, to affect calcineurin signaling and CRTC-1-dependent transcription ( Mallick et al, 2021b ). Recently, we identified several genes ( cpz-1/CTSZ , cdk-1/CDK1 , rnr-1/RRM1 , his-7/H2AX , and ard-1/HSD17B10 ) that function downstream of pry-1 to regulate the lifespan and stress response of animals ( Mallick et al, 2021a ). Taken together, these findings demonstrate that pry-1 acts as a master regulator of aging-related processes and functions by coordinating the activities of diverse genes and pathways.…”

“…RNAi-mediated gene silencing was performed using a protocol previously published by our laboratory ( Mallick et al, 2021a ). Plates were seeded with Escherichia coli HT115 expressing either dsRNA specific to candidate genes or empty vector (L4440).…”

“…Synchronized gravid adults were bleached, and eggs were plated. After becoming young adults, animals were analyzed for stress sensitivity and lifespan ( Mallick et al, 2021a ).…”

“…Thus, it is essential to identify and understand the function of pathway components so that they can be targeted not only to promote normal growth, but also to develop potential treatments for associated diseases. To this end, our group is investigating the function of an Axin scaffolding protein homolog in the nematode Caenorhabditis elegans , PRY-1, which is necessary for the formation of many tissues and cell types, as well as for maintaining stress responses and aging ( Mallick et al, 2019b , 2020 , 2021a ; Mallick and Gupta, 2022 ). To gain insights into the genetic network of pry-1, we performed mRNA and miRNA transcriptome profiling experiments ( Ranawade et al, 2018 ; Mallick et al, 2019a ).…”