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Dissecting the Subnuclear Localization Patterns of Argonaute Proteins and Other Components of the RNA-Directed DNA Methylation Pathway in Plants

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Plant Argonaute Proteins

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1640))

Abstract

RNA-directed DNA methylation (RdDM) is a nuclear pathway which is comprised of multiple main and accessory protein components, including two plant-specific DNA-dependent RNA polymerases, Pol IV and Pol V, and argonaute (AGO) proteins. Regulation in the RdDM pathway can be achieved via multiple mechanisms, including the spatial distribution of different RdDM components. Here we describe a protocol for dissecting the subnuclear localization of AGO proteins and other RdDM components, including nuclei extraction from seedlings, slide preparation, and subsequent immunostaining.

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Acknowledgments

This work was supported by the Chinese Academy of Sciences and by National Institutes of Health Grant R01GM070795 (to J.–K. Z).

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Authors and Affiliations

  1. Shanghai Center for Plant Stress Biology and Center of Excellence in Molecular Plant Sciences, Chinese Academy of Sciences, 320 Yueyang Road, Shanghai, 200031, China

    Cheng-Guo Duan & Jian-Kang Zhu

  2. Department of Horticulture and Landscape Architecture, Purdue University, West Lafayette, IN, USA

    Cheng-Guo Duan & Jian-Kang Zhu

Authors
  1. Cheng-Guo Duan
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  2. Jian-Kang Zhu
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Corresponding author

Correspondence to Cheng-Guo Duan .

Editor information

Editors and Affiliations

  1. Instituto de Biología Molecular y Celular de Plantas, (Consejo Superior de Investigaciones Científicas–Universidad Politécnica de Valencia), Valencia, Spain

    Alberto Carbonell

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Duan, CG., Zhu, JK. (2017). Dissecting the Subnuclear Localization Patterns of Argonaute Proteins and Other Components of the RNA-Directed DNA Methylation Pathway in Plants. In: Carbonell, A. (eds) Plant Argonaute Proteins. Methods in Molecular Biology, vol 1640. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7165-7_8

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  • DOI: https://doi.org/10.1007/978-1-4939-7165-7_8

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-7164-0

  • Online ISBN: 978-1-4939-7165-7

  • eBook Packages: Springer Protocols

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