Oxygen-independent stabilization of HIF-2a in breast cancer through direct interaction with peptidyl-prolyl cis-trans isomerase NIMA-interacting 1

Abstract

Peptidyl-prolyl cis-trans isomerase NIMA-interacting 1 (Pin1) isomerizes the nearby proline (Pro) residue when it detects phosphorylated serine (Ser) or threonine (Thr) of target proteins, altering their structure, stability, function, and interaction with other proteins. Hypoxia-inducible factor 2a (HIF-2a), a transcription factor that transactivates many oncogenic genes under hypoxic conditions, harbours the pSer/Thr-Pro motif. We found for the first time that Pin1 binds to HIF-2a physically in normoxic as well as hypoxic conditions in human breast cancer cells. The level of ubiquitinated HIF-2a was significantly raised by Pin1 knockdown, while expression of its mRNA transcript was unaffected. In agreement with this observation, the cycloheximide chase assay demonstrated that Pin1 prolonged the stability of HIF-2a. Serine 672, 696, and 790 of HIF-2a were found to undergo phosphorylation. Of these, the main amino acid involved in the Pin1 binding and HIF-2a stabilization was identified as serine 790, located in the nuclear export signal region of HIF-2a. The tissue array with human breast cancer specimens showed elevated expression of HIF-2a as well as Pin1 compared to adjacent normal tissues. Knockdown of Pin1 or HIF-2a diminished breast cancer cell migration and colony formation. In conclusion, Pin1 stabilizes HIF-2a through direct interaction, which contributes to the growth of breast cancer.