FBXL5 modulates HIF-1α transcriptional activity by degradation of CITED2

• FBXL5 interacts with CITED2 and promotes its degradation by the proteasome.
• FBXL5 interferes with CITED2 interaction with p300 and CBP in the cells.
• Overexpression of FBXL5 in mouse embryonic stem cells reduces Cited2 protein levels.
• FBXL5-mediated degradation of CITED2 leads to the activation of HIF-1α.

CITED2 is a ubiquitously expressed nuclear protein exhibiting a high affinity for the cysteine–histidine–rich domain 1 (CH1) of the transcriptional co-activators CBP/p300. CITED2 is particularly efficient in the inhibition of the hypoxia-inducible factor-1α (HIF-1α) dependent transcription by competing with it for the interaction with the CH1 domain. Here we report a direct and specific interaction between CITED2 and the F-box and leucine rich repeat protein 5 (FBXL5), a substrate adaptor protein which is part of E3 ubiquitin ligase complexes mediating protein degradation by the proteasome. We demonstrated that depletion of FBXL5 by RNA interference led to an increase of CITED2 protein levels. Conversely, overexpression of FBXL5 caused the decrease of CITED2 protein levels in a proteasome-dependent manner, and impaired the interaction between CITED2 and the CH1 domain of p300 in living cells. In undifferentiated mouse embryonic stem cells, the overexpression of FBXL5 also reduced Cited2 protein levels. Finally, we evidenced that FBXL5 overexpression and the consequent degradation of CITED2 enabled the transcriptional activity of the N-terminal transactivation domain of HIF-1α. Collectively, our results highlighted a novel molecular interaction between CITED2 and FBXL5, which might regulate the steady state CITED2 protein levels and contribute to the modulation of gene expression by HIF-1α.