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Fig. 3. The oskar 3'UTR is sufficient to rescue the oogenesis arrest phenotype of oskar RNA null alleles. (A,C) Schematic of oskar alleles and transgenes. Solid black bars represent the oskar promoter and 5'UTR (left) and the 3'UTR (right). M1 and M2 are the two translation initiation sites of oskar. E1 through E4 indicate the oskar exons. Black dots in osk⁵⁴, osk⁸⁴, osk³⁴⁶ and P(mM1 mM2stop) transcripts show the positions of the stop codons in the nonsense alleles and the transgene. P(osk+) is a wild-type oskar transgene that encodes both the long and short Oskar isoforms and fully rescues the oskar strong loss of function alleles (Markussen et al., 1995). Black dots at M1 and M2 in P(oskM1L), P(oskM139L), P(mM1 mM2stop) and P(mM1SLmM2) indicate the mutated translation initiation sites. HP7 represents the hairpin loop sequence (Kozak, 1989a) inserted between M1 and M2, and the grey dot shows the frame-shift mutation inserted in P(mM1SLmM2). (C) Light grey bars represent the UAS promoter. UAS oskWT expresses the oskar gene (including introns) under the control of the yeast UAS promoter. UAS oski(1,2,3) expresses an oskar RNA whose three introns have been deleted (dark grey bar), under control of the UAS promoter. Note that all UAS-driven transcripts contain an intron derived from the pUASp vector. Dashed line in UAS osk-K10 indicates the K10 3'UTR. UAS osk3'UTR expresses only the 3'UTR of oskar without any oskar coding region under control of UAS. To ensure continued high-level expression throughout oogenesis, expression of all UAS transgenes was driven by pCOG:Gal4VP16 and nanos:Gal4VP16 simultaneously. Flies were grown at 25^oC. In the case of the UAS osk-K10 transgene, rescue was also tested at 29^oC, at which Gal4-induced expression is maximal. (B,D) Average number and standard deviation produced daily by females of the genotype indicated underneath (normalized to the average number of eggs laid by wild-type females). See text for details. (B) Grey bar: Oregon-R control. Black and white bars: alleles or transgenes in osk^A87/Df(3R)p^XT103 and osk¹⁸⁷/Df(3R)p^XT103 background, respectively. (D) Grey bar: w¹¹¹⁸ control. Black bars: transgenes in osk^A87/Df(3R)p^XT103 background.

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