Knockout of cyclin dependent kinases 8 and 19 leads to depletion of cyclin C and suppresses spermatogenesis and male fertility in mice

Abstract

AbstractParalogs CDK8 and CDK19 are regulatory kinases associated with the transcriptional Mediator complex. We have for the first time generated mice with the systemic inducibleCdk8knockout on the background ofCdk19constitutive knockout.Cdk8/19double knockout (DKO) males, but not singleCdk8andCdk19KO, had an atrophic reproductive system and were infertile. The DKO males lacked postmeiotic spermatids and spermatocytes after meiosis I pachytene. Testosterone levels were decreased whereas the amounts of the luteinizing hormone were unchanged. Single cell RNA sequencing showed marked differences in the expression of steroidogenic genes (such asCyp17a1, StarandFads) in Leydig cells concomitant with alterations in Sertoli cells and spermatocytes likely associated with impaired synthesis of steroids.StarandFadswere also downregulated in cultivated Leydig cells after DKO. The treatment of primary Leydig cells culture with a CDK8/19 inhibitor did not induce the same changes in gene expression as DKO, and prolonged treatment of mice with a CDK8/19 inhibitor did not affect the size of testes. DKO, in contrast to single knockouts or treatment with a CDK8/19 kinase inhibitor, led to depletion of cyclin C (CcnC), the binding partner of CDK8/19 that has been implicated in CDK8/19-independent functions. This suggests that the observed phenotype was likely mediated through kinase-independent activities of CDK8/19, such as CcnC stabilization.