Mutated SPOP E3 ligase promotes 17βHSD4 protein degradation to drive androgenesis and prostate cancer progression.

Molecular mechanisms underlying intratumoral androgenesis and aberrant androgen receptor (AR) activation in prostate cancer (PCa) remain poorly understood. Here we demonstrate that ectopic expression of the E3 ubiquitin ligase adaptor speckle-type poxvirus and zinc finger domain protein (SPOP) stabilizes 17βHSD4. SPOP bound a functional substrate-binding consensus (SBC) motif 315RATST319 in 17βHSD4 and promoted non-degradable K27- and K29-linked poly-ubiquitination of 17βHSD4. The effect of SPOP was antagonized by serum- and glucocorticoid kinase-3 (SGK3)-mediated phosphorylation of serine 318 (S318) in the SBC and S318 phosphorylation-dependent binding of SKP2 E3 ligase and subsequent K48-linked poly-ubiquitination and proteasomal degradation of 17βHSD4. PCa-associated SPOP mutations impaired the SPOP-17βHSD4 interaction, caused 17βHSD4 protein destruction in PCa cells in culture and patient specimens, and increased testosterone production and PCa cell growth in vitro and in mouse models. Thus, we have identified SPOP and SKP2 as two essential E3 ubiquitin ligases that exert opposite effects on 17βHSD4 protein degradation and intratumoral androgenesis in PCa cells. We further demonstrate that SPOP mutations or SKP2 overexpression contribute to PCa progression by decreasing 17βHSD4 expression and increasing intratumoral androgen synthesis.