CCR4-NOT subunit CCF-1/CNOT7 promotes transcriptional activation to multiple stress responses in Caenorhabditis elegans.

CCR4-NOT is a versatile eukaryotic protein complex that controls multiple steps in gene expression regulation from synthesis to decay. In yeast, CCR4-NOT has been implicated in stress response regulation, though this function in other organisms remains unclear. In a genome-wide RNAi screen, we identified a subunit of the CCR4-NOT complex, ccf-1, as a requirement for the C. elegans transcriptional response to cadmium and acrylamide stress. Using whole-transcriptome RNA sequencing, we show that the knockdown of ccf-1 attenuates the activation of a broad range of stress-protective genes in response to cadmium and acrylamide, including those encoding heat shock proteins and xenobiotic detoxification. Consistently, survival assays show that the knockdown of ccf-1 decreases C. elegans stress resistance and normal lifespan. A yeast 2-hybrid screen using a CCF-1 bait identified the homeobox transcription factor PAL-1 as a physical interactor. Knockdown of pal-1 inhibits the activation of ccf-1 dependent stress genes and reduces C. elegans stress resistance. Gene expression analysis reveals that knockdown of ccf-1 and pal-1 attenuates the activation of elt-2 and elt-3 under stress that encode master transcriptional co-regulators of stress response in the C. elegans, and that overexpression of ELT-2 can suppress ccf-1's requirement for gene transcription in a stress-dependent manner. Our findings reveal a new role for CCR4-NOT in the environmental stress response and define its role in stress resistance and longevity in C. elegans.

Neuron-specific toxicity of chronic acrylamide exposure in C. elegans.

Acrylamide is a food-borne chemical with well-known neurotoxic properties. To date, the toxicity mechanisms of chronic acrylamide exposure are not fully understood. Using the genetic model Caenorhabditis elegans, we found that chronic acrylamide exposure induces a locomotor defect that is characterized by severe uncoordination of muscle movement that is distinct from an overall reduction in activity. C. elegans exhibiting chronic acrylamide-induced locomotor defects show significant degeneration to the dopaminergic and cholinergic, but not GABAergic motor neurons. Degeneration of the dopaminergic and cholinergic neurons are found in 58% to 67% of C. elegans after chronic acrylamide exposure, with the varying degrees of severity ranging from neuronal blebbing to complete dendrite loss. The observed pattern of neurotoxicity does not have a heritable effect, as parental exposure to chronic acrylamide does not lead to neurodegeneration in the developed offspring. Overall, these finding illustrate that chronic acrylamide exposure cause locomotor defects by inducing degeneration of specific neuron types in C. elegans.