The mouse protein phosphatase gene Ppp1cc is essential for male fertility, with mutants displaying a failure in spermatogenesis including a widespread loss of post-meiotic germ cells and abnormalities in the mitochondrial sheath. This phenotype is hypothesized to be attributed to the loss of the testis specific isoform PPP1CC2. To identify PPP1CC2 interacting proteins with a function in spermatogenesis, we performed GST pull down assays in mouse testis lysate. Amongst the identified candidate interactors was the testis specific protein kinase TSSK1, which is also essential for male fertility. Subsequent interaction experiments have confirmed the capability of PPP1CC2 to form a complex with TSSK1 mediated by direct interaction of each with the kinase substrate protein TSKS. Interaction between PPP1CC2 and TSKS is mediated through an RVxF docking motif on the TSKS surface. Phosphoproteomic analysis of the mouse testis identified a novel serine phosphorylation site within the TSKS RVxF motif that appears to negatively regulate binding to PPP1CC2. Immunohistochemical analysis of TSSK1 and TSKS in Ppp1cc mutant testis showed reduced accumulation to distinct cytoplasmic foci and other abnormalities in their distribution consistent with the loss of germ cells and seminiferous tubule disorganization observed in the Ppp1cc mutant phenotype. A comparison of Ppp1cc and Tssk1/2 knockout phenotypes via electron microscopy revealed similar abnormalities in the morphology of the mitochondrial sheath. This data demonstrates a novel kinase/phosphatase complex in the testis that could play a critical role in the completion of spermatogenesis.