Letter | Published:

LIN28 phosphorylation by MAPK/ERK couples signalling to the post-transcriptional control of pluripotency

Nature Cell Biology volume 19, pages 6067 (2017) | Download Citation

Abstract

Signalling and post-transcriptional gene control are both critical for the regulation of pluripotency1,2, yet how they are integrated to influence cell identity remains poorly understood. LIN28 (also known as LIN28A), a highly conserved RNA-binding protein, has emerged as a central post-transcriptional regulator of cell fate through blockade of let-7 microRNA biogenesis and direct modulation of mRNA translation3. Here we show that LIN28 is phosphorylated by MAPK/ERK in pluripotent stem cells, which increases its levels via post-translational stabilization. LIN28 phosphorylation had little impact on let-7 but enhanced the effect of LIN28 on its direct mRNA targets, revealing a mechanism that uncouples LIN28’s let-7-dependent and -independent activities. We have linked this mechanism to the induction of pluripotency by somatic cell reprogramming and the transition from naive to primed pluripotency. Collectively, our findings indicate that MAPK/ERK directly impacts LIN28, defining an axis that connects signalling, post-transcriptional gene control, and cell fate regulation.

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Acknowledgements

We thank P. Sharp, L. Cantley, G. Ruvkun, and members of the Daley laboratory for invaluable discussions, A. D. L. Angeles for critical feedback on the manuscript, X. Wu/Yi Zhang’s laboratory and R. Rubio/DFCI CCCB for assistance with RNA-seq, and R. Tomaino at the Taplin Biological Mass Spectrometry Core for assistance with mass spectrometry. Bioanalyser analysis was performed in the BCH IDDRC Molecular Genetics Core, which is supported by NIH (NIH-P30-HD 18655). Sequencing analysis was conducted on the Orchestra High Performance Computing Cluster at Harvard Medical School. K.M.T. was an HHMI International Student Research Fellow and a Herchel Smith Graduate Fellow. D.S.P. was supported by a grant from NIGMS (T32GM007753). R.I.G. was supported by a grant from NIGMS (R01GM086386). G.Q.D. is an investigator of the Howard Hughes Medical Institute and the Manton Center for Orphan Disease Research, and was supported by a grant from NIGMS (R01GM107536).

Author information

Affiliations

  1. Stem Cell Transplantation Program, Division of Hematology/Oncology, Manton Center for Orphan Disease Research, Boston Children’s Hospital and Dana-Farber Cancer Institute, Boston, Massachusetts 02115, USA

    • Kaloyan M. Tsanov
    • , Daniel S. Pearson
    • , Zhaoting Wu
    • , Areum Han
    • , Marc T. Seligson
    • , John T. Powers
    • , Jihan K. Osborne
    •  & George Q. Daley
  2. Department of Biological Chemistry and Molecular Pharmacology, Harvard Stem Cell Institute, Harvard Medical School, Boston, Massachusetts 02115, USA

    • Kaloyan M. Tsanov
    • , Daniel S. Pearson
    • , Zhaoting Wu
    • , Areum Han
    • , Robinson Triboulet
    • , Marc T. Seligson
    • , John T. Powers
    • , Jihan K. Osborne
    • , Richard I. Gregory
    •  & George Q. Daley
  3. Howard Hughes Medical Institute, Boston, Massachusetts 02115, USA

    • Kaloyan M. Tsanov
    • , Daniel S. Pearson
    • , Zhaoting Wu
    • , Areum Han
    • , Marc T. Seligson
    • , John T. Powers
    • , Jihan K. Osborne
    •  & George Q. Daley
  4. Stem Cell Program, Division of Hematology/Oncology, Boston Children’s Hospital, Boston, Massachusetts 02115, USA

    • Robinson Triboulet
    •  & Richard I. Gregory
  5. Cell Signaling Technology, Inc., Danvers, Massachusetts 01923, USA

    • Susan Kane
  6. Department of Cell Biology, Harvard Medical School, Boston, Massachusetts 02115, USA

    • Steven P. Gygi

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Contributions

K.M.T. designed and performed the experiments, and wrote the manuscript. D.S.P. helped with RNA-seq, HeLa clonal series generation and expression analysis. Z.W. performed reprogramming experiments. A.H. performed RNA-seq bioinformatics analysis. R.T. and R.I.G. shared unpublished results and generated the isogenic HeLa cells. M.T.S. performed expression analysis. J.T.P. and J.K.O. helped with experimental design. S.K. generated the human pLIN28A antibody. S.P.G. supervised the proteomics experiments. G.Q.D. designed and supervised experiments, and wrote the manuscript.

Competing interests

G.Q.D. and R.I.G. hold options and intellectual property related to 28/7 Therapeutics, a company seeking to develop inhibitors of the LIN28/let-7 pathway. S.K. is an employee of Cell Signaling Technology.

Corresponding author

Correspondence to George Q. Daley.

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DOI

https://doi.org/10.1038/ncb3453

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