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LIN-28 co-transcriptionally binds primary let-7 to regulate miRNA maturation in Caenorhabditis elegans

Nature Structural & Molecular Biology volume 18, pages 302–308 (2011)Cite this article

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Abstract

The highly conserved let-7 microRNA (miRNA) regulates developmental pathways across animal phyla. Mis-expression of let-7 causes lethality in C. elegans and has been associated with several human diseases. We show that timing of let-7 expression in developing worms is under complex transcriptional and post-transcriptional control. Expression of let-7 primary transcripts oscillates during each larval stage, but precursor and mature let-7 miRNAs do not accumulate until later in development after LIN-28 protein has diminished. We demonstrate that LIN-28 binds endogenous primary let-7 transcripts co-transcriptionally. We further show that LIN-28 binds endogenous primary let-7 transcripts in the nuclear compartment of human ES cells, suggesting that this LIN-28 activity is conserved across species. We conclude that co-transcriptional interaction of LIN-28 with let-7 primary transcripts blocks Drosha processing and, thus, precocious expression of mature let-7 during early development.

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Figure 1: Expression of let-7 is transcriptionally and post-transcriptionally regulated.
Figure 2: Developmentally regulated processing of let-7 pri-miRNA transcripts.
Figure 3: Regulation of let-7 processing by LIN-28.
Figure 4: LIN-28 binds endogenous let-7 primary transcripts in C. elegans and human ES cells.
Figure 5: LIN-28 binds endogenous let-7 genomic DNA.

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Acknowledgements

We thank J. Lykke-Andersen (Univ. California, San Diego (UCSD)) and members of the Pasquinelli laboratory for their suggestions and critical reading of this manuscript. We thank E. Moss (Univ. Medicine and Dentistry of New Jersey) for providing the plin-28:LIN-28:GFP strain, R. Gassmann and A. Desai (UCSD) for providing the polyclonal antibody to GFP, M. Li and M. David (UCSD) for sharing their real-time PCR machine, and the Caenorhabditis Genetics Center for worm strains. P.M.V.W. was supported by a Ruth L. Kirschstein National Research Service Award (F32GM087004) from the US National Institute of General Medical Sciences. Z.S.K. was supported by a US National Institutes of Health (NIH) Cellular and Molecular Genetics and NIH/National Cancer Institute T32 CA009523 Graduate Student Training Grants. V.H.B. was supported by a UCSD Chancellor's Undergraduate Research Scholarship. This work was funded by the NIH (GM071654), the Keck, Searle, V., Emerald and Peter Gruber Foundations (A.E.P.) and by the California Institute of Regenerative Medicine (RB1-01413) and the Stem Cell Program at UCSD (G.W.Y.).

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Authors and Affiliations

  1. Department of Biology, University of California, San Diego, La Jolla, California, USA

    Priscilla M Van Wynsberghe, Zoya S Kai, Victoria H Burton & Amy E Pasquinelli

  2. Department of Cellular and Molecular Medicine, University of California, San Diego, La Jolla, California, USA

    Katlin B Massirer & Gene W Yeo

  3. Stem Cell Program, University of California, San Diego, La Jolla, California, USA

    Katlin B Massirer & Gene W Yeo

  4. Institute for Genomic Medicine, University of California, San Diego, La Jolla, California, USA

    Katlin B Massirer & Gene W Yeo

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  1. Priscilla M Van Wynsberghe
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Contributions

A.E.P. and P.M.V. designed the project and wrote the paper; P.M.V. (all figures), Z.S.K. (Fig. 1 and Supplementary Fig. 1), K.B.M. (Fig. 4) and V.H.B. (Fig. 4) performed the experiments; A.E.P. and G.W.Y. supervised the studies.

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Correspondence to Amy E Pasquinelli.

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The authors declare no competing financial interests.

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Supplementary Figures 1–7, Supplementary Tables 1–4 and Supplementary Methods (PDF 1257 kb)

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Van Wynsberghe, P., Kai, Z., Massirer, K. et al. LIN-28 co-transcriptionally binds primary let-7 to regulate miRNA maturation in Caenorhabditis elegans. Nat Struct Mol Biol 18, 302–308 (2011). https://doi.org/10.1038/nsmb.1986

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