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An atypical RNA polymerase involved in RNA silencing shares small subunits with RNA polymerase II

Nature Structural & Molecular Biology volume 16, pages 9193 (2009) | Download Citation

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Abstract

Genetic evidence indicates that plant-specific homologs of DNA-dependent RNA polymerase (Pol) II large subunits form Pol IV and Pol V complexes involved in small interfering RNA production and RNA-directed DNA methylation. Here we describe evidence that Pol V contains subunits shared with Pol II, but that RNA polymerase II subunit (RPB)-4 is missing from Pol V and that RPB5 is present as a Pol V–specific isomer, RPB5b. Pol V also has other proteins that are not present in Pol II, consistent with a role of this complex as an effector of silencing.

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Acknowledgements

We thank T. Guilfoyle from the University of Missouri-Columbia for providing plant Pol II antibodies and helpful discussions. We thank M. Mann from the Max-Planck-Institut in Martinsried for hosting A.M.E.J. for the analysis of samples S4 and S5. We also thank Arabidopsis T-DNA mutant providers: The Salk Institute, GABI-Kat, the Wisconsin pDS/Lox Project and the Nottingham Arabidopsis Stock Centre. L.H. was partly supported by an Overseas Research Students award and a Marie Curie EST Grant (504273). The Gatsby Charitable Foundation provided funding for this work.

Author information

Affiliations

  1. The Sainsbury Laboratory, John Innes Centre, Colney Lane, Norwich NR4 7UH, UK.

    • Linfeng Huang
    • , Alexandra M E Jones
    •  & David C Baulcombe
  2. Department for Proteomics and Signal Transduction, Max-Planck Institute for Biochemistry, Am Klopferspitz 18, D-82152 Martinsried, Germany.

    • Nina C Hubner
  3. Department of Plant Sciences, University of Cambridge, Downing Street, Cambridge CB2 3EA, UK.

    • Linfeng Huang
    • , Iain Searle
    • , Kanu Patel
    • , Hannes Vogler
    •  & David C Baulcombe

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Contributions

D.C.B. and L.H. designed the experiments; L.H., A.M.E.J. and N.C.H. performed immunoprecipitation and MS experiments; K.P., I.S., H.V. and L.H. characterized mutant plants; D.C.B., L.H., A.M.E.J. and I.S. wrote the paper.

Corresponding author

Correspondence to David C Baulcombe.

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    Supplementary Text and Figures

    Supplementary Figures 1–11, Supplementary Tables 1–6 and Supplementary Methods

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DOI

https://doi.org/10.1038/nsmb.1539

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