Unknown,Transcriptomics,Genomics,Proteomics

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Gene expression in Drosophila Poly mutant and wild-type

ABSTRACT: The Poly mutant was identified as a previously uncharacterised, but essential gene in Drosphila. Loss of Poly reults in lethality at the third larval instar stage, but only after a stage of extreme larval longevity. We aimed to assess the consequences of loss of the Poly gene on glocal gene expression by microarray analysis. Metabolism is altered in Poly mutants and here we find altered gene expression in insulin signaling pathways. Four biological replicates each of Poly mutant and wild-type larvae were used to assess gene expression on FL002 microarrays (GPL5135) . These data reveal that Poly is a mediatior of insulin receptor/TOR signaling. Four biological replicates of poly mutant and wild-type Drosophila. Two arrays had wild-type in Cy5 channel and poly in Cy3 channel, the other two were reversed.

ORGANISM(S): Drosophila melanogaster

SUBMITTER: Donald Dunbar

PROVIDER: E-GEOD-32637 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Project description:Analysis of differential gene expression in third instar Drosophila salivary glands in the absence versus presence of cohesin. ABSTRACT: Developmental abnormalities observed in Cornelia de Lange Syndrome (CdLS) have been genetically linked to mutations in the cohesin machinery. These findings raise the possibility that cohesin, in addition to its canonical function of mediating sister chromatid cohesion, might also be involved in regulating gene expression. We report that cleavage of cohesinM-CM-^Us kleisin subunit in post-mitotic Drosophila salivary glands induces major changes (both up and down) in the transcript levels of many genes. Kinetic analyses of changes in transcript levels upon cohesin cleavage reveal that a subset of genes responds to cohesin cleavage within a few hours. In addition, cohesin binds to most of these loci, suggesting that cohesin is directly regulating their expression. Amongst these genes are several that are regulated by the steroid hormone Ecdysone. Transcripts at EcR and Eip74EF, which encode an Ecdysone Receptor and an Ecdysone-regulated transcription factor, respectively, decline ten-fold within four hours of cohesin cleavage. Cytological visualization of transcription at selected Ecdysone-responsive genes reveals that puffing at Eip74EF ceases within an hour or two of cohesin cleavage, long before any decline in EcR associated with this locus. We conclude that cohesin regulates expression of a distinct set of genes, including those mediating the Ecdysone response. A heat-inducible transgene (hs-TEV) was used to induce TEV in terminally differentiated third instar Drosophila salivary glands expressing either wild type (+ cohesin) or TEV-cleavable myc10-tagged Rad21 protein (Rad21TEV, - cohesin). Total RNA was isolated from + and - cohesin salivary glands 10-12 hours after heat shock induction of TEV (7 independent biological samples each). RNA samples were converted to cDNA, labeled with Cy3 and Cy5 respectively (3x) and vice versa (4x; dye swaps), and hybridized to INDAC FL003 arrays. Analysis of seven arrays, each hybridized to an independently generated sample-pair revealed major differences in transcript levels between + and - cohesin samples.

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Gene expression in Drosophila Poly mutant and wild-type

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