Unknown,Transcriptomics,Genomics,Proteomics

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HEXIM knockdown triggers apoptosis-induced proliferation and deregulates Hedgehog signaling

ABSTRACT: We address the function of HEXIM, an inhibitor of the general transcriptional elongation regulator P-TEFb which regulates the transcriptional status of many developmental genes, during Drosophila development. We showed that HEXIM knockdown mutants display organs development failure. In the wing disc, it induces apoptosis and affects Hh signaling. The continuous death of proliferative cells is compensated by apoptosis-induced cell proliferation, in a manner similar to that of differentiated cells, together with high levels of Hh and Ci. We completed this analysis with microarrays to characterize the molecular phenotype of HEXIM knockdown during eye differentiation. HEXIM knockdown during eye differentiation was carried out with a UAS-Hexim-RNAi strain crossed with a GMR-Gal4 driver strain (Bloomington stock center)). Adult GMR>RNAi Hexim flies display slightly smaller, rough and often necrosed eyes. Total RNA were collected from isolated heads. We then used microarray to compare the transcriptome of HEXIM knockdown mutant with that of wild type flies.

ORGANISM(S): Drosophila melanogaster

SUBMITTER: Patricia Uguen

PROVIDER: E-GEOD-54590 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Project description:Purpose: Combination of systemic RNAi with RNAseq to identify target genes of important pathways in early development Drosophila early patterning occurs in the syncytial blastoderm where transcription factors diffuse between cells. However, in typical insect embryos, patterning occurs in a cellularized environment where signaling pathways are likely to play a more fundamental role. We use the short germ beetle Tribolium castaneum to investigate two putative Wnt and Hh signaling centers located in the anlagen of head and growth zone. These structures are known to develop in a different way in short germ insects. We find that Hh acts upstream of Wnt in the head, which is different from the Drosophila situation. In the growth zone Wnt signaling acts upstream. For the first time outside Drosophila, we comprehensively determine the Wnt and Hh target gene sets and distinguish the anterior from the posterior gene sets by genetically depleting head or growth zone. Surprisingly, there are significantly more targets in the growth zone than in the head for both pathways and we find that their growth zone gene sets are essentially non-overlapping. Furthermore, several pair rule genes, Tc-caudal, Tc-twist and hindgut patterning genes are regulated by Wnt signaling. In addition, Wnt controls growth zone metabolism and cell division. Posterior Hh signaling activates several genes potentially involved in a proteinase cascade of unknown function. Unexpectedly, we find the Wnt target Tc-senseless to be required for hindgut development. 10-11 h old whole embyro mRNA profiles of the following treatments: reference: wild type 100bp single read triplicates reference: wild type 50bp single read triplicates treatment: Tc-arrow RNAi knockdown 50 bp single read triplicates treatment: Tc-frizzled1/2 RNAi double knockdown 50 bp single read triplicates treatment: Tc-hedgehog RNAi knockdown 100bp single reads quadruplicates treatment: Tc-orthodenticle RNAi knockdown 100bp single reads triplicates treatment: Tc-torso RNAi knockdown 100bp single reads triplicates treatment: Tc-wntless RNAi knockdown 50bp single reads triplicates Total: 2 references, 6 treatments, 25 sequencing runs

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HEXIM knockdown triggers apoptosis-induced proliferation and deregulates Hedgehog signaling

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