Dp71 is regulated by phosphorylation and ubiquitin-proteasome system in neuronal cells.

The Dystrophin (Dp) gene is responsible for Duchenne muscular dystrophy (DMD), which is characterized by progressive muscular degeneration and variable degrees of cognitive impairment. Although Dp71 is the most abundant among the Dp isoforms in the brain, the regulatory mechanisms of the related expression levels have not been elucidated. In this study, we found that the constitutive expression levels of Dp71 in PC12 cells were sensitive to proteasomal inhibition. The ectopic expression of FLAG-tagged ubiquitin revealed that Dp71 was ubiquitinated intracellularly. Interestingly, proteasomal inhibition was accompanied by a posttranslational accumulation of modified Dp71, which was restored by protein phosphatase treatment in vitro, indicating that phosphorylation is responsible for the modification and affects the proteasome-dependent degradation of Dp71. Proteasomal activity-sensitive phosphorylated Dp71 is closely associated with syntrophin, a well-known binding partner of Dp71, and syntrophin is also regulated by proteasomal activity in a similar way to Dp71, suggesting that the posttranslational regulatory machinery for Dp71 level is coupled with Dp71-syntrophin molecular complex. Taken together, our results indicated that the expression levels of Dp71 are posttranslationally regulated by the phosphorylation-ubiquitin-proteasomal pathway, which may indicate the presence of regulatory mechanisms underlying the proteostasis of both Dp and its molecular complex, which may lead to better therapeutic approaches for the treatment of Dp-related diseases.