Skip to main content

Thank you for visiting You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

EGFR modulates microRNA maturation in response to hypoxia through phosphorylation of AGO2


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.

This is a preview of subscription content, access via your institution

Relevant articles

Open Access articles citing this article.

Access options

Buy article

Get time limited or full article access on ReadCube.


All prices are NET prices.

Figure 1: EGFR interacts with AGO2 in response to hypoxia.
Figure 2: EGFR modulates miRNA maturation in response to hypoxia.
Figure 3: EGFR phosphorylates AGO2 at Tyr 393 to suppress the maturation of long-loop mHESM in response to hypoxia.
Figure 4: p-Y393-AGO2 enhances cell survival and invasiveness under hypoxia and correlates with poorer overall survival in breast cancer patients.


  1. Kim, V. N. MicroRNA biogenesis: coordinated cropping and dicing. Nature Rev. Mol. Cell Biol. 6, 376–385 (2005)

    Article  CAS  Google Scholar 

  2. van Kouwenhove, M., Kedde, M. & Agami, R. MicroRNA regulation by RNA-binding proteins and its implications for cancer. Nature Rev. Cancer 11, 644–656 (2011)

    Article  CAS  Google Scholar 

  3. Lu, J. et al. MicroRNA expression profiles classify human cancers. Nature 435, 834–838 (2005)

    Article  ADS  CAS  Google Scholar 

  4. Leung, A. K. & Sharp, P. A. MicroRNA functions in stress responses. Mol. Cell 40, 205–215 (2010)

    Article  CAS  Google Scholar 

  5. Pouysségur, J., Dayan, F. & Mazure, N. M. Hypoxia signalling in cancer and approaches to enforce tumour regression. Nature 441, 437–443 (2006)

    Article  ADS  Google Scholar 

  6. Gould, G. W. & Lippincott-Schwartz, J. New roles for endosomes: from vesicular carriers to multi-purpose platforms. Nature Rev. Mol. Cell Biol. 10, 287–292 (2009)

    Article  CAS  Google Scholar 

  7. Mosesson, Y., Mills, G. B. & Yarden, Y. Derailed endocytosis: an emerging feature of cancer. Nature Rev. Cancer 8, 835–850 (2008)

    Article  CAS  Google Scholar 

  8. Cikaluk, D. E. et al. GERp95, a membrane-associated protein that belongs to a family of proteins involved in stem cell differentiation. Mol. Biol. Cell 10, 3357–3372 (1999)

    Article  CAS  Google Scholar 

  9. Eulalio, A., Huntzinger, E. & Izaurralde, E. Getting to the root of miRNA-mediated gene silencing. Cell 132, 9–14 (2008)

    Article  CAS  Google Scholar 

  10. Diederichs, S. & Haber, D. A. Dual role for argonautes in microRNA processing and posttranscriptional regulation of microRNA expression. Cell 131, 1097–1108 (2007)

    Article  CAS  Google Scholar 

  11. Chendrimada, T. P. et al. TRBP recruits the Dicer complex to Ago2 for microRNA processing and gene silencing. Nature 436, 740–744 (2005)

    Article  ADS  CAS  Google Scholar 

  12. Lemmon, M. A. & Schlessinger, J. Cell signaling by receptor tyrosine kinases. Cell 141, 1117–1134 (2010)

    Article  CAS  Google Scholar 

  13. Wang, Y. et al. Regulation of endocytosis via the oxygen-sensing pathway. Nature Med. 15, 319–324 (2009)

    Article  CAS  Google Scholar 

  14. Franovic, A. et al. Translational up-regulation of the EGFR by tumor hypoxia provides a nonmutational explanation for its overexpression in human cancer. Proc. Natl Acad. Sci. USA 104, 13092–13097 (2007)

    Article  ADS  CAS  Google Scholar 

  15. Reynolds, A. R., Tischer, C., Verveer, P. J., Rocks, O. & Bastiaens, P. I. EGFR activation coupled to inhibition of tyrosine phosphatases causes lateral signal propagation. Nature Cell Biol. 5, 447–453 (2003)

    Article  CAS  Google Scholar 

  16. Lee, O. H. et al. Genome-wide YFP fluorescence complementation screen identifies new regulators for telomere signaling in human cells. Mol. Cell. Proteom. 10, M110.001628 (2010)

    Article  Google Scholar 

  17. Jiang, X., Huang, F., Marusyk, A. & Sorkin, A. Grb2 regulates internalization of EGF receptors through clathrin-coated pits. Mol. Biol. Cell 14, 858–870 (2003)

    Article  CAS  Google Scholar 

  18. Bertout, J. A., Patel, S. A. & Simon, M. C. The impact of O2 availability on human cancer. Nature Rev. Cancer 8, 967–975 (2008)

    Article  CAS  Google Scholar 

  19. Ventura, A. & Jacks, T. MicroRNAs and cancer: short RNAs go a long way. Cell 136, 586–591 (2009)

    Article  CAS  Google Scholar 

  20. Nicoloso, M. S., Spizzo, R., Shimizu, M., Rossi, S. & Calin, G. A. MicroRNAs – the micro steering wheel of tumour metastases. Nature Rev. Cancer 9, 293–302 (2009)

    Article  CAS  Google Scholar 

  21. Leung, A. K. & Sharp, P. A. MicroRNAs: a safeguard against turmoil? Cell 130, 581–585 (2007)

    Article  CAS  Google Scholar 

  22. Schirle, N. T. & MacRae, I. J. The crystal structure of human Argonaute2. Science 336, 1037–1040 (2012)

    Article  ADS  CAS  Google Scholar 

  23. Elkayam, E. et al. The structure of human Argonaute-2 in complex with miR-20a. Cell 150, 100–110 (2012)

    Article  CAS  Google Scholar 

  24. Tahbaz, N. et al. Characterization of the interactions between mammalian PAZ PIWI domain proteins and Dicer. EMBO Rep. 5, 189–194 (2004)

    Article  CAS  Google Scholar 

  25. Maniataki, E. & Mourelatos, Z. A human, ATP-independent, RISC assembly machine fueled by pre-miRNA. Genes Dev. 19, 2979–2990 (2005)

    Article  CAS  Google Scholar 

  26. Tsutsumi, A., Kawamata, T., Izumi, N., Seitz, H. & Tomari, Y. Recognition of the pre-miRNA structure by Drosophila Dicer-1. Nature Struct. Mol. Biol. 18, 1153–1158 (2011)

    Article  CAS  Google Scholar 

  27. Suzuki, H. I. et al. Modulation of microRNA processing by p53. Nature 460, 529–533 (2009)

    Article  ADS  CAS  Google Scholar 

Download references


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).

Author information

Authors and Affiliations



J.S. and M.-C.H. designed and conceived the study; J.S. and M.-C.H. wrote the manuscript; J.L.H. contributed to the preparation of the manuscript. J.S., W.X., Y.B.K., L.H., S.-O.L., Y.D., Y. Wang, W.-C.C. and C.-H.C. did the experiments; Y. Wu provided human primary breast tumour samples; Y.C.L. provided the split-half-YFP-fused constructs; X.L. and C.-G.L. assisted in next-generation RNA deep sequencing; B.P.J. provided the pipeline analysis service for RNA sequencing data; and K.N. and D.-J.P. analysed the crystal structure of human AGO2.

Corresponding author

Correspondence to Mien-Chie Hung.

Ethics declarations

Competing interests

The authors declare no competing financial interests.

Supplementary information

Supplementary Information

This file contains Supplementary Figures 1–41, Supplementary Table 1, Supplementary Methods and Supplementary References. (PDF 15146 kb)

Supplementary Data

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)

PowerPoint slides

Rights and permissions

Reprints and Permissions

About this article

Cite this article

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).

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI:

This article is cited by


By submitting a comment you agree to abide by our Terms and Community Guidelines. If you find something abusive or that does not comply with our terms or guidelines please flag it as inappropriate.


Quick links

Nature Briefing

Sign up for the Nature Briefing newsletter — what matters in science, free to your inbox daily.

Get the most important science stories of the day, free in your inbox. Sign up for Nature Briefing