A role for the Perlman syndrome exonuclease Dis3l2 in the Lin28–let-7 pathway

Abstract

The pluripotency factor Lin28 blocks the expression of let-7 microRNAs in undifferentiated cells during development, and functions as an oncogene in a subset of cancers1. Lin28 binds to let-7 precursor (pre-let-7) RNAs and recruits 3′ terminal uridylyl transferases to selectively inhibit let-7 biogenesis2,3,4. Uridylated pre-let-7 is refractory to processing by Dicer, and is rapidly degraded by an unknown RNase5. Here we identify Dis3l2 as the 3′–5′ exonuclease responsible for the decay of uridylated pre-let-7 in mouse embryonic stem cells. Biochemical reconstitution assays show that 3′ oligouridylation stimulates Dis3l2 activity in vitro, and knockdown of Dis3l2 in mouse embryonic stem cells leads to the stabilization of pre-let-7. Our study establishes 3′ oligouridylation as an RNA decay signal for Dis3l2, and identifies the first physiological RNA substrate of this new exonuclease, which is mutated in the Perlman syndrome of fetal overgrowth and causes a predisposition to Wilms’ tumour development6.

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Figure 1: Dis3l2 is associated with uridylated pre-let-7.
Figure 2: Dis3l2 preferentially degrades uridylated pre-let-7.
Figure 3: Molecular determinants of Dis3l2 activity.
Figure 4: Dis3l2 is required for degradation of uridylated pre-let-7 in ESCs.

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Acknowledgements

We thank R. LaPierre for technical assistance and F.-L. Meng for Rrp44 and Exosc10 shRNA. Thanks to the Children’s Hospital Boston/IDDRC Proteomics Core for mass spectrometry. R.I.G. was supported by grants from the US National Institute of General Medical Sciences (NIGMS) (R01GM086386) and The American Cancer Society (121635-RSG-11-175-01-RMC). J.E.T. was supported by a pre-doctoral fellowship from the National Science Foundation. R.T. was supported by the Wolbach fellowship from Boston Children’s Hospital.

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H.-M.C. designed and performed most of the experiments in Figs 1 and 2, and all of the experiments in Fig. 4. H.-M.C., R.T. and J.E.T. designed and performed experiments in Figs 1, 2, 3, as well as the Supplementary Figs. R.I.G. and H.-M.C. wrote the paper with input from R.T. and J.E.T.

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Correspondence to Richard I. Gregory.

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The authors declare no competing financial interests.

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This file contains Supplementary Figures 1-9 and Supplementary Methods. (PDF 1476 kb)

Supplementary Data

This file contains raw mass spectrometry data. (XLSX 641 kb)

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Chang, H., Triboulet, R., Thornton, J. et al. A role for the Perlman syndrome exonuclease Dis3l2 in the Lin28–let-7 pathway. Nature 497, 244–248 (2013). https://doi.org/10.1038/nature12119

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