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. 2016 Apr 25;113(19):5269–5274. doi: 10.1073/pnas.1600936113

Fig. 3.

Fig. 3.

Disruption of the normal PSI-U1 snRNP interaction changes the AS of gene transcripts functionally enriched for male courtship behavioral regulation, including fruitless, the master courtship regulator gene. (A) Flowchart of the procedure used to compare the transcriptome-wide AS patterns in male fly heads from the PSIΔAB fly strain versus full-length PSI. (B) Summary of identified differentially spliced gene transcripts in PSIΔAB male fly heads versus full-length PSI strain by three independent computational methods, MISO (12) (green circle), JuncBase (13) (yellow circle), and JUM (14) (blue circle). See also Fig. S2 and Datasets S1–S3. (C) GO analysis of the AS target gene transcripts of PSIΔAB. Categories of related genes are listed at the left and enrichment significance (−log FDR) is indicated along the x axis. (D) The PSIΔAB mutant affects the AS of the three male-specific fruitless mRNA isoforms that are functionally distinct for maintaining normal male courtship behavior (cruciality fru-C > fru-B > fru-A) (17). Here the 5′ and 3′ ends of the fruitless pre-mRNA are shown. Green rectangles represent exons and black lines introns. The male-specific fruitless mRNA isoforms use an alternative 3′SS in the second exon and thus bypass a premature stop codon (shown by the stop sign) that is exclusively included in female fruitless isoforms, resulting in only male-specific transcripts encoding functional proteins. RNA-seq tracks (34) from PSIΔAB and full-length PSI male fly heads are shown below. Arcs represent the splice junctions specific for fru-A, fru-B, and fru-C, respectively, with the number of reads mapped to each junction shown. The overall expression of fruitless stays the same for PSIΔAB and wild-type samples, as shown by the fragments per kilobase per million (FPKM) value. (E) RT-qPCR verification of the AS of fru-A, -B, and -C in PSIΔAB male fly head (pink) versus full-length PSI (blue). RNA level of fru-A normalized to rp49 (y axis on the left) and level of fru-B and fru-C normalized to fru-A (y axis on the right). Mean of male head RNA samples from three separate PSIΔAB or full-length PSI fly lines with ±SE is shown. Significance was analyzed by one-way ANOVA test. n.s., not significant; **P value < 0.02, statistically significant.