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Summary: |
Regulatory Factor X (RFX) transcription factors (TF) are present in most eukaryotes and are important for regulating processes such as the cell cycle, brain development, neural function and ciliogenesis. C. elegans RFX TF, DAF-19, has four known isoforms: A, B, C, and M. Recent studies suggest DAF-19 A/B, the longest isoforms varying only by one exon, are important in synaptic protein maintenance. However, the molecular details of this regulation are not yet understood. It may be activating genes coding for proteins that inhibit synaptic protein degradation or inhibiting genes coding for proteins that activate synaptic protein degradation. Failing to maintain healthy synapses have been shown to correlate with neurodegenerative diseases such as, Alzheimer’s. The purpose of this study was to identify the gene targets of DAF-19A/B in C. elegans. Transgenes for three putative genes, hex-1, T07F10.1, and Y73F8A.11, were constructed so that the promoter regions of each gene regulate the expression of GFP. Each transgene was injected into two strains of animals: one with a functional daf-19 gene and one with a daf-19 mutation. The expression of GFP was compared between these two strains, via confocal microscopy, to identify if DAF-19 regulates the expression of that gene. All genes showed DAF-19-dependent changes in their expressions. Furthermore, behavioral investigation was carried out, via a roaming and dwelling assay, to observe if the mutants of these putative genes phenocopy DAF-19 mutants. Mutating either of these three genes caused a daf-19-mutant like behavior. Finally, a rescue experiment was carried out by injecting a translational fusion of the genes of interest with GFP.
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