Project description:This study was designed to identify changes in gene expression resulting from depletion of Mago nashi (Mago), a core subunit of the exon junction complex. Drosophila S2R+ cells were treated with double-stranded RNA targeting either LacZ or Mago for 6 days. Poly(A)+ RNA was purified from each sample and sequenced using 54 bp reads on an Illumina Genome Analyzer II. Fold changes in expression were calculated for each gene as the ratio of the reads per kilobase per million reads (RPKM) for Mago relative to LacZ. In other experiments, we have shown that Mago is required for correct splicing of the MAP kinase gene and its depletion causes a large reduction in MAP kinase mRNA levels. MAP kinase is a large gene in heterochromatin. In this study, we find that heterochromatic genes are disproportionately down-regulated in Mago-depleted cells, and those with large introns are particularly likely to be affected. SRA accession number: SRP003001.1 Sequencing of poly(A)+ RNA from S2R+ cells treated with either lacZ (control) or mago (experimental) dsRNA
Project description:This study was designed to identify changes in gene expression resulting from depletion of Mago nashi (Mago), a core subunit of the exon junction complex. Drosophila S2R+ cells were treated with double-stranded RNA targeting either LacZ or Mago for 6 days. Poly(A)+ RNA was purified from each sample and sequenced using 54 bp reads on an Illumina Genome Analyzer II. Fold changes in expression were calculated for each gene as the ratio of the reads per kilobase per million reads (RPKM) for Mago relative to LacZ. In other experiments, we have shown that Mago is required for correct splicing of the MAP kinase gene and its depletion causes a large reduction in MAP kinase mRNA levels. MAP kinase is a large gene in heterochromatin. In this study, we find that heterochromatic genes are disproportionately down-regulated in Mago-depleted cells, and those with large introns are particularly likely to be affected. SRA accession number: SRP003001.1
Project description:We have studied the deoxynucleotide transport in Drosophila melanogaster. On the basis of homology with the S. cerevisiae RIM2 gene, encoding a pyrimidine deoxynucleotide carrier (Marrobbio et al. 2006), the CG18317 gene (dRIM2) in the fruit fly may code for a deoxynucleotide carrier. We demonstrated that Drosophila S2R+ cells, silenced for the dRIM2 expression, had a marked defect in the amounts of all mitochondrial dNTPs, both purines and pyrimidines. In vivo dRIM2 homozygous knockout produced a larval lethal phenotype. dRIM2-/- larvae showed (i) impairments in the locomotor behavior, (ii) a decrease in the rates of oxygen consumption and (iii) a depletion of the mtDNA. Following a detailed morphological characterization carried out in dRIM2-/- larvae evidenced an ongoing mitochondrial biogenesis accompanied by an alteration of mitochondria shaping. Additionally, the role of dRIM2 in the purine and pyrimidine metabolism was supported by a microarray analysis. We conclude that dRIM2 is a Drosophila deoxynucleotide carrier, essential for maintaining the mitochondrial functionality.