Identification of nuclear and cytoplasmic neuronal mRNA targets of fly orthologues of the human motor neuron disease proteins Smn, Fus and TDP-43

This study aims to identify the neuronal nuclear and cytoplasmic mRNA targets of the drosophila orthologues of the human Spinal Muscular Atrophy (Smn) and Amyotrophic Lateral Sclerosis (Fus and TDP-43) disease proteins. GFP or Smn, caz or TBPH GFP-fusion proteins were expressed conditionally in the adult nervous system via the Gal4/UAS system. Lysates were prepared from heads and fractionated into cytoplasmic and nuclear fractions. Nuclear proteins were further solubilized with high salt buffer and recovered in the nuclear soluble fraction. The cytoplasmic and nuclear soluble fractions were used for immunoprecipitation with GFP-trap beads. Co-immunoprecipitated RNAs from 2-5 independent replicate assays were purified and subjected to Illumina paired-end mRNA-seq.