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. 2019 Jan 29;15(1):e1007899. doi: 10.1371/journal.pgen.1007899

Fig 5. The role of TCTP and CSN4 in the control of CULLIN neddylation and cell proliferation is conserved in Drosophila.

Fig 5

(a) Downregulation of dTCTP specifically in the eyes (ey>dTCTPi) of Drosophila leads to small eye phenotype. Close up images of wild type adult eye and of eye expressing ey>dTCTPi are shown (Bars = 500 μm). (b) Drosophila adult retina visualized by immersion microscopy with the Tomato/GFP-FLP/FRT method. CSN4 mutant clones are visualized by the expression of Rh1-GFP and by the absence of tomato (red) and are delineated by a white line. Photoreceptors are visualized by Rh1-GFP (Left panels), Rh1-tdTomato (Middle panels) or in the merge (Right panels). Photoreceptor absence is indicated by loss or diffuse GFP fluorescence in CSN4k08018 mutant clones (CSN4-/-, upper panels). CSN4k08018 mutant clones in which p35 is overexpressed (CSN4-/-, lower panels) still show loss or diffuse GFP staining in mutant clone. Bars = 5μm. (c-e) CSN4 mutant photoreceptors show a delay in the acquisition of neuronal identity but not in caspase staining. Third instar eye imaginal discs carrying dCSN4-/- mutant clones visualized by the lack of GFP (green) Bars = 25 μm. (c) eye imaginal discs stained with anti-Dcp-1 (purple) show no increase of staining in dCSN4-/- mutant clones. (d) Eye imaginal discs stained with anti-Elav show a delay of expression (arrowheads) at the morphogenetic furrow and a reduced level in dCSN4-/- mutant clones. (e) Quantification of Elav staining in (d) shows significant reduction in dCSN4-/- mutant clones compared to wild type (GFP) (n = 5; p < 0,01). (f) Downregulation of dTCTP in all tissues of Drosophila larvae (tub>dTCTPi) leads to developmental arrest with reduced size and subsequent larval lethality at the first instar larvae. 7 days after egg-laying, control larvae (tub-GAL4) are at third instar (left) while tub>dTCTPi larvae stay at first instar (right). Bars = 500 μm. (g) Impaired larval development is associated with a decrease of the CUL1NEDD8 abundance in the tub>dTCTPi compared to tub-GAL4. Star: free CUL1; Arrow: CUL1NEDD8 form.