The RNA-binding Proteins FMR1, Rasputin and Caprin Act Together with the UBA Protein Lingerer to Restrict Tissue Growth in Drosophila melanogaster
Figure 2
Eyes overexpressing lig are reduced in size and are partially rescued by CycE and DIAP1 co-overexpression.
(A–I) Scanning electron micrographs of eyes overexpressing the indicated UAS transgenes under the control of ey-Gal4 during development from flies reared on 25% (A–C), 100% (D–F) and 400% (G–I) yeast-containing food, respectively. Scale bar represents 100 µm. (J) Statistical analyses as described in Figure 1D: ey>GFP (741±30 and 753±12), ey>lig (628±31; p = 1.66E-05 and 608±14; p = 1.17E-10) and ey>ligR185C/UTR (448±80; p = 4.35E-06 and 435±54; p = 8.12E-09) eyes at 25% yeast-containing food, ey>GFP (768±14 and 768±12), ey>lig (666±30; p = 3.03E-05 and 626±39; p = 3.09E-05) and ey>ligR185C/UTR (287±42; p = 9.50E-09 and 306±62; p = 5.38E-07) eyes at 100% yeast-containing food and ey>GFP (748±34 and 785±21), ey>lig (588±35; p = 1.55E-06 and 619±53; p = 6.71E-05) and ey>ligR185C/UTR (193±50; p = 1.18E-10 and 253±36; p = 2.84E-11) eyes at 400% yeast-containing food. (K–L) lig overexpressing clones (induced with the Actin-Flp out-Gal4 system and marked by GFP) in wing imaginal discs of third instar larvae undergo apoptosis as judged by Cleaved Caspase-3 staining (red) (L–L') in comparison to the control (K–K'). Scale bar represents 50 µm. (M–R) The reduced size of eyes overexpressing ligR185C/UTR induced by ey-Gal4 (N) is partially rescued by co-overexpression of DIAP1, an inhibitor of apoptosis (P), or CycE (R). Overexpression of DIAP1 (O) or CycE (Q) has no effect on eye size in comparison to the control (M). Scale bar represents 100 µm. (S) Statistical analyses as described in Figure 1D: ey>GFP (770±15 and 746±14), ey>ligR185C/UTR (309±34 and 135±73), ey>GFP, DIAP1 (789±17 and 790±16), ey>ligR185C/UTR, DIAP1 (477±47; p = 9.60E-06 and 410±51; p = 7.97E-06), ey>GFP, CycE (753±33 and 708±42), and ey>ligR185C/UTR, CycE (446±38; p = 1.47E-05 and 488±46; p = 1.01E-07) eyes. (T–U) The rescue of the reduced size of eyes overexpressing ligR185C/UTR induced by ey-Gal4 is improved by co-overexpression of DIAP1 and CycE (U) in comparison to the rescue experiment of co-overexpression of DIAP1 or CycE. Co-overexpression of DIAP1 and CycE has no effect on eye size in a wild-type background (T). Scale bar represents 100 µm. (V) Statistical analyses as described in Figure 1D: ey>ligR185C/UTR (201±83 and 275±35), ey>CycE, DIAP1 (744±14 and 712±37), ey>ligR185C/UTR, CycE, DIAP1 (551±23; p = 1.45E-05 and 595±29; p = 5.38E-10) Genotypes: (A, D and G) w/y w; ey-Gal4/UAS-GFP (B, E and H) w/y w; ey-Gal4/+; UAS-lig/+ (C, F, and I) w/y w; ey-Gal4/+; UAS-ligR185C/UTR/+ (K) y w hsFLP/y w; UAS-GFP/+; Act>CD2>Gal4, UAS-GFP/+ (L) y w hsFLP/y w; Act>CD2>Gal4, UAS-GFP/UAS-lig [86Fb] (M–R) first experiment: (M) w/y w; ey-Gal4/UAS-GFP (N) w/y w; ey-Gal4/+; UAS-ligR185C/UTR/+ (O) w/y w; ey-Gal4/UAS-GFP; EP-DIAP1/+ (P) w/y w; ey-Gal4/+; UAS-ligR185C/UTR/EP-DIAP1 (Q) w/y w; ey-Gal4/UAS-GFP; UAS-CycE/+ (R) w/y w; ey-Gal4/+; UAS-ligR185C/UTR/UAS-CycE; second experiment: (M) w/y w; ey-Gal4/UAS-GFP; UAS-lacZ (N) w/y w; ey-Gal4/UAS-GFP; UAS-ligR185C/UTR/+ (O) w/y w; ey-Gal4/UAS-GFP; EP-DIAP1/+ (P) w/y w; ey-Gal4/+; UAS-ligR185C/UTR/EP-DIAP1 (Q) w/y w; ey-Gal4/UAS-GFP; UAS-CycE/+ (R) w/y w; ey-Gal4/+; UAS-ligR185C/UTR/UAS-CycE (T) w/y w; ey-Gal4/UAS-CycE; EP-DIAP1/+ (U) w/y w; ey-Gal4/UAS-CycE; UAS-ligR185C/UTR/EP-DIAP1.