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

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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Figure 1: Co-immunoprecipitation and western blot to confirm shared Pol II and Pol V subunits.
Figure 2: Molecular phenotype of RPB5b and KTF1 mutants.

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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.

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Authors

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.

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Correspondence to David C Baulcombe.

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Supplementary Figures 1–11, Supplementary Tables 1–6 and Supplementary Methods (PDF 5969 kb)

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Huang, L., Jones, A., Searle, I. et al. An atypical RNA polymerase involved in RNA silencing shares small subunits with RNA polymerase II. Nat Struct Mol Biol 16, 91–93 (2009). https://doi.org/10.1038/nsmb.1539

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