The testis-specific proteasome subunit Pros6T of D. melanogaster is required for individualization and nuclear maturation during spermatogenesis

Most regulated proteolysis in eukaryotes is carried out by the 26S proteasome. This large, multisubunit complex comprises a catalytic core particle (20S proteasome) and a regulatory particle (19S regulator) capping each end. In Drosophila, about a third of the 32 proteasome subunits are found to have testis-specific isoforms, encoded by paralogous genes. Here, we characterize in detail the spermatogenic expression of the core particle subunit Pros6 (Pros35) and its testis-specific isoform Pros6T. Using GFP-tagged transgenes, it is shown that whereas the Pros6 subunit is expressed in early stages of spermatogenesis, gradually fading away following meiosis, the testis-specific Pros6T becomes prominent in spermatid nuclei and cytoplasm after meiosis, and persists in mature sperm. In addition, these subunits are found in numerous 'speckles' near individualization complexes, similar to the previously described expression pattern of the caspase Dronc (Nedd2-like caspase), suggesting a link to the apoptosis pathway. We also studied the phenotypes of a loss-of-function mutant of Pros6T generated by targeted homologous recombination. Homozygous males are sterile and show spermatogenic defects in sperm individualization and nuclear maturation, consistent with the expression pattern of Pros6T. The results demonstrate a functional role of testis-specific proteasomes during Drosophila spermatogenesis.