Project description:This study aims to elucidate how the core circadian transcription factor, CLOCK (Clk), mediates diet-dependent transcriptional changes in fly heads. In particular, we sought to identify how CLK influences gene-level expression changes in fly heads from flies that had been reared on either a high nutrient or low nutrient (dietary restriction) diet.
Project description:CLK targets from fly heads using the TIM-GAL4; UAS-CLKGR line Experiment Overall Design: Fly heads were stimulated with vehicle + CHX or dexamethasone + CHX
Project description:To identify the eye-enriched genes by comparing the mRNA expression profiles from wild type fly heads (CantonS) and eye-less fly heads (sine occulis).
Project description:We applied ribosomal footprinting experiment followed by RNA-seq in order to detect circular RNA associated with ribosomes in fly heads.
Project description:To address the contribution of transcriptional regulation to Drosophila clock gene expression and to behavior, we generated a series of CRISPR-mediated deletions within two regions of the circadian gene timeless (tim), an intronic E box region and an upstream E box region that are both recognized by the key transcription factor Clock (Clk) and its heterodimeric partner Cycle. The upstream deletions but not an intronic deletion dramatically impact tim expression in fly heads; the biggest upstream deletion reduces peak RNA levels and tim RNA cycling amplitude to about 15% of normal, and there are similar effects on tim protein (TIM). The cycling amplitude of other clock genes is also strongly reduced, in these cases due to increases in trough levels. To examine the effect of promoter E-box deletions on Clk binding, we performed Clk-ChIP around the clock in wild-type and 126 mutant flies.
Project description:Influence of diet and neuronal clk (clock) activity on hemolymph proteomics. We have shown that as photoreceptors die (in the fly) they necrose, which results in their intercellular contents leaking into the hemolymph. We hypothesize that this process is regulated by diet and circadian clock control.
Analysis of differential protein expression in the hemolymph from flies reared on a high protein diet. Comparison of flies with and without a functional circadian clock within their photoreceptors.
Species/Strain: Drosophila, Elav-GeneSwitch-GAL4>UAS-Clk-DN1 (+/- RU486), female