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SMN and symmetric arginine dimethylation of RNA polymerase II C-terminal domain control termination

Nature volume 529, pages 4853 (07 January 2016) | Download Citation

Abstract

The carboxy-terminal domain (CTD) of the RNA polymerase II (RNAP II) subunit POLR2A is a platform for modifications specifying the recruitment of factors that regulate transcription, mRNA processing, and chromatin remodelling. Here we show that a CTD arginine residue (R1810 in human) that is conserved across vertebrates is symmetrically dimethylated (me2s). This R1810me2s modification requires protein arginine methyltransferase 5 (PRMT5) and recruits the Tudor domain of the survival of motor neuron (SMN, also known as GEMIN1) protein, which is mutated in spinal muscular atrophy. SMN interacts with senataxin, which is sometimes mutated in ataxia oculomotor apraxia type 2 and amyotrophic lateral sclerosis. Because POLR2A R1810me2s and SMN, like senataxin, are required for resolving RNA–DNA hybrids created by RNA polymerase II that form R-loops in transcription termination regions, we propose that R1810me2s, SMN, and senataxin are components of an R-loop resolution pathway. Defects in this pathway can influence transcription termination and may contribute to neurodegenerative disorders.

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Primary accessions

Gene Expression Omnibus

Data deposits

Data for ChIP-seq analyses have been deposited at GEO with the accession code GSE73379.

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Acknowledgements

We thank J. Manley for GST–CTD constructs; D. Reinberg for RNAP II me2a antibodies; D. Eick for wild-type and RNAP II (R1810A) constructs; and S. Leppla for purified S9.6 antibodies. We also thank D. Torti and D. Leung for Illumina library preparation and sequencing, T. Hajian for the purified PRMT5–WDR77 complex, J. Li for purified 8WG16 antibodies, and D. Durocher for constructive criticism and advice during the course of this work. This project was supported by the Ontario Research Fund from the Ontario Ministry of Research and Innovation (to J.F.G. and T.P.) and by CIHR Operating Grants to J.F.G. and B.J.B. D.Y.Z. was supported by a National Science and Engineering Research Council of Canada Studentship and an Ontario Graduate Scholarship. U.B. was supported by a long-term Postdoctoral Fellowship from HFSP. B.J.B. holds the University of Toronto Banbury Chair in Medical Research.

Author information

Author notes

    • Gerald Gish
    •  & Ulrich Braunschweig

    These authors contributed equally to this work.

Affiliations

  1. Donnelly Centre, University of Toronto, Toronto, Ontario M5S 3E1, Canada.

    • Dorothy Yanling Zhao
    • , Ulrich Braunschweig
    • , Yue Li
    • , Zuyao Ni
    • , Frank W. Schmitges
    • , Guoqing Zhong
    • , Jason Moffat
    • , Benjamin J. Blencowe
    •  & Jack F. Greenblatt
  2. Lunenfeld-Tanenbaum Research Institute, Mount Sinai Hospital, 600 University Avenue, Toronto, Ontario M5G 1X5, Canada

    • Dorothy Yanling Zhao
    • , Gerald Gish
    •  & Tony J. Pawson
  3. Department of Molecular Genetics, University of Toronto, Toronto, Ontario M5S 1A8, Canada

    • Dorothy Yanling Zhao
    • , Jason Moffat
    • , Tony J. Pawson
    • , Benjamin J. Blencowe
    •  & Jack F. Greenblatt
  4. Department of Computer Science, University of Toronto, Toronto, Ontario M5S 3G4, Canada

    • Yue Li
  5. Structural Genomics Consortium, University of Toronto, Toronto, Ontario M5G 1L7, Canada

    • Ke Liu
    • , Weiguo Li
    • , Masoud Vedadi
    •  & Jinrong Min

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Contributions

J.F.G. supervised the project. D.Y.Z. performed the experiments. J.F.G. and D.Y.Z. wrote the manuscript. T.J.P., B.J.B., Z.N., and G.G. commented on experiments and edited the manuscript. G.G. prepared the FITC peptides. F.W.S. performed ChIP-seq experiments. U.B. and Y.L. performed computational data analysis for ChIP-seq. Vectors for shRNAs were provided by J.M., and G.Z. generated stable shRNA-mediated knockdown cell lines. Z.N. generated CRISPR knockout cell lines. J.M. and K.L. provided the purified Tudor domains. M.V. provided the purified PRMT5–WDR77 complex. W.L. performed isothermal titration calorimetry assays.

Competing interests

The authors declare no competing financial interests.

Corresponding author

Correspondence to Jack F. Greenblatt.

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    Supplementary Information

    This file contains Supplementary Table 1 which shows the number of times each type of experiment described in this work was performed with the reported results) and raw western blots used in the paper.

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DOI

https://doi.org/10.1038/nature16469

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