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The  Drosophila  polypyrimidine tract-binding protein (dmPTB or  hephaestus ) plays an important role during spermatogenesis. The  heph   2   mutation in this gene results in a specific defect in spermatogenesis, causing aberrant spermatid individualization and male sterility. However, the array of molecular defects in the mutant remains uncharacterized. Using an unbiased high throughput sequencing approach, we have identified transcripts that are misregulated in this mutant. Aberrant transcripts show altered expression levels, exon skipping, and alternative 5’ ends. We independently verified these findings by reverse-transcription and polymerase chain reaction (RT-PCR) analysis. Our analysis shows misregulation of transcripts that have been connected to spermatogenesis, including components of the actomyosin cytoskeletal apparatus. We show, for example, that the  Myosin light chain 1  ( Mlc1)  transcript is aberrantly spliced. Furthermore, bioinformatics analysis reveals that  Mlc1  contains a high affinity binding site(s) for dmPTB and that the site is conserved in many  Drosophila  species. We discuss that  Mlc1  and other components of the actomyosin cytoskeletal apparatus offer important molecular links between the loss of dmPTB function and the observed developmental defect in spermatogenesis. This study provides the first comprehensive list of genes misregulated  in vivo  in the  heph   2   mutant in  Drosophila  and offers insight into the role of dmPTB during spermatogenesis.

plos one

High Throughput Sequencing Identifies Misregulated Genes in the Drosophila Polypyrimidine Tract-Binding Protein (hephaestus) Mutant Defective in Spermatogenesis