The APC/C E3 Ligase Complex Activator FZR1 Restricts BRAF Oncogenic Function

Abstract

BRAF drives tumorigenesis by coordinating the activation of the RAS/RAF/MEK/ERK oncogenic signaling cascade. However, upstream pathways governing BRAF kinase activity and protein stability remain undefined. Here, we report that in primary cells with active APC^FZR1, APC^FZR1 earmarks BRAF for ubiquitination-mediated proteolysis, whereas in cancer cells with APC-free FZR1, FZR1 suppresses BRAF through disrupting BRAF dimerization. Moreover, we identified FZR1 as a direct target of ERK and CYCLIN D1/CDK4 kinases. Phosphorylation of FZR1 inhibits APC^FZR1, leading to elevation of a cohort of oncogenic APC^FZR1 substrates to facilitate melanomagenesis. Importantly, CDK4 and/or BRAF/MEK inhibitors restore APC^FZR1 E3 ligase activity, which might be critical for their clinical effects. Furthermore, <i>FZR1</i> depletion cooperates with AKT hyperactivation to transform primary melanocytes, whereas genetic ablation of <i>Fzr1</i> synergizes with <i>Pten</i> loss, leading to aberrant coactivation of BRAF/ERK and AKT signaling in mice. Our findings therefore reveal a reciprocal suppression mechanism between FZR1 and BRAF in controlling tumorigenesis.<b>Significance:</b> FZR1 inhibits BRAF oncogenic functions via both APC-dependent proteolysis and APC-independent disruption of BRAF dimers, whereas hyperactivated ERK and CDK4 reciprocally suppress APC^FZR1 E3 ligase activity. Aberrancies in this newly defined signaling network might account for BRAF hyperactivation in human cancers, suggesting that targeting CYCLIN D1/CDK4, alone or in combination with BRAF/MEK inhibition, can be an effective anti-melanoma therapy. <i>Cancer Discov; 7(4); 424-41. ©2017 AACR.</i><i>See related commentary by Zhang and Bollag, p. 356</i><i>This article is highlighted in the In This Issue feature, p. 339</i>.