Epigenetics of ER-hormesis in dietary restricted C. elegans
Identifying epigenetic regulators of proteostasis longevity in the Endoplasmic reticulum conferred by dietary restriction.
Project background: This project was started as a continuation of the research done at the molecular aging laboratory at National Institute of Immunology, New Delhi. The lab had previously shown that the dietary restriction model eat-2(-) C. elegans worms had better unfolded protein responses (UPR) of the endoplasmic reticulum (ER) in adulthood, an increased health-span and lifespan because of a transient protein folding stress during larval development (L2 stage). However, the mechanism behind this hormetic response remained elusive. I hypothesized that the transient UPR stress induced a cellular memory during larval development which was epigenetic in nature. Therefore, I planned to conduct an RNA-interference screen that would knock down established epigenetic regulators in C. elegans and observe for the presence or absence of the same hormetic response in the eat-2(-) worms.
Project importance: The mechanisms through which dietary restriction promotes longevity have been investigated in much detail. However, this study showed that dietary restriction possibly establishes an epigenetic change upstream of proteostasis pathways in the ER which is maintained till adulthood to provide increased resistance against protein folding stresses.
Skills learned:
- C. elegans handling, bleaching, crossing, stack preparation
- NGM media preparation
- Bacterial culture (primary, secondary), C. elegans feed preparation
- On-plate Dietary restriction protocols (peptone restriction, bacterial/solid dilution)
- Tunicamycin- UPR assay (induced UPR)
- PCR, Gel electrophoresis, Western Blotting
- Fluorescence microscopy (GFP-tagged hsp-4 chaperone for observing UPR activation).
