MicroRNAs (miRNAs) are generated by two-step processing to yield small RNAs that negatively regulate target gene expression at the post-transcriptional level1. Deregulation of miRNAs has been linked to diverse pathological processes, including cancer2,3. Recent studies have also implicated miRNAs in the regulation of cellular response to a spectrum of stresses4, such as hypoxia, which is frequently encountered in the poorly angiogenic core of a solid tumour5. However, the upstream regulators of miRNA biogenesis machineries remain obscure, raising the question of how tumour cells efficiently coordinate and impose specificity on miRNA expression and function in response to stresses. Here we show that epidermal growth factor receptor (EGFR), which is the product of a well-characterized oncogene in human cancers, suppresses the maturation of specific tumour-suppressor-like miRNAs in response to hypoxic stress through phosphorylation of argonaute 2 (AGO2) at Tyr 393. The association between EGFR and AGO2 is enhanced by hypoxia, leading to elevated AGO2-Y393 phosphorylation, which in turn reduces the binding of Dicer to AGO2 and inhibits miRNA processing from precursor miRNAs to mature miRNAs. We also identify a long-loop structure in precursor miRNAs as a critical regulatory element in phospho-Y393-AGO2-mediated miRNA maturation. Furthermore, AGO2-Y393 phosphorylation mediates EGFR-enhanced cell survival and invasiveness under hypoxia, and correlates with poorer overall survival in breast cancer patients. Our study reveals a previously unrecognized function of EGFR in miRNA maturation and demonstrates how EGFR is likely to function as a regulator of AGO2 through novel post-translational modification. These findings suggest that modulation of miRNA biogenesis is important for stress response in tumour cells and has potential clinical implications.
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We thank B. Pickering, D. Yu, and A.-B. Shyu for suggestions and technical assistance with northern blot analysis. This work was supported by the US National Institutes of Health (CA109311 and CA099031 to M.-C.H., and CCSG Core Grant CA16672), the US National Breast Cancer Foundation, The Center for Biological Pathway at the UT MD Anderson Cancer Center, S. G. Komen (SAC110016 to M.-C.H.), The Sister Institution Fund of China Medical University and Hospital and the UT MD Anderson Cancer Center, the Cancer Research Center of Excellence (D0H102-TD-C-111-005, Taiwan), a Private University grant (NSC99-2632-B-039-001-MY3, Taiwan), and the Program for Stem Cell and Regenerative Medicine Frontier Research (NSC101-2321-B-039-001, Taiwan).
The authors declare no competing financial interests.
This file contains Supplementary Figures 1–41, Supplementary Table 1, Supplementary Methods and Supplementary References. (PDF 15146 kb)
This file contains the Normalized Expression (RPKM) of mRNAs that are regulated by EGFR and likely to be targeted by Top-Scoring mHESM. (XLS 68 kb)
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Shen, J., Xia, W., Khotskaya, Y. et al. EGFR modulates microRNA maturation in response to hypoxia through phosphorylation of AGO2. Nature 497, 383–387 (2013). https://doi.org/10.1038/nature12080
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