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Characterization of Plant Polyadenylation Complexes by Using Tandem Affinity Purification

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Polyadenylation in Plants

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1255))

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Abstract

Messenger RNA in eukaryotic cells is initially produced as a nascent transcript (pre-mRNA) without a polyadenine [poly(A)] tail to the 3′ end. The precise cleavage of the pre-mRNA and addition of a poly(A) track need the communication between cis-elements in the pre-mRNA sequences and transacting protein factors recognizing them. Based on homology analyses, Arabidopsis cleavage and polyadenylation specificity factor (AtCPSF) complex should play a critical role in pre-mRNA 3′ end processing. Here we describe the isolation of AtCPSF complex by using a tandem affinity purification (TAP) method. We demonstrate that TAP is a potent protein complex isolating approach that can fulfill a downstream protein identification purpose based on mass spectrometry techniques.

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Acknowledgement

We received funding support from the US National Science Foundation (grant nos. IOS–0817829 and IOS-1353354 to QQL). QQL received funding support from the Fujian Hundred Talent Plan.

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Authors and Affiliations

  1. Department of Plant Pathology, College of Plant Protection, Nanjing Agricultural University, Nanjing, 210095, China

    Hongwei Zhao

  2. Institute of Fruit Trees, Fujian Academy of Agricultural Science, Fuzhou, 350013, China

    Xinfu Ye

  3. Rice Research Institute, Fujian Academy of Agricultural Science, Fuzhou, 350013, China

    Qingshun Quinn Li

  4. Department of Biology, Miami University, Oxford, OH, 45056, USA

    Qingshun Quinn Li

Authors
  1. Hongwei Zhao
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  2. Xinfu Ye
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  3. Qingshun Quinn Li
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Corresponding author

Correspondence to Hongwei Zhao .

Editor information

Editors and Affiliations

  1. Department of Plant and Soil Sciences, University of Kentucky, Lexington, Kentucky, USA

    Arthur G. Hunt

  2. Department of Biology, Key Laboratory of the Ministry of Education for Coastal and Wetland Ecosystems, College of the Environment and Ecology, Miami University, Oxford, OH, USA and Xiamen University, Xiamen, Fujian, Fujian, China

    Qingshun Quinn Li

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Zhao, H., Ye, X., Li, Q.Q. (2015). Characterization of Plant Polyadenylation Complexes by Using Tandem Affinity Purification. In: Hunt, A., Li, Q. (eds) Polyadenylation in Plants. Methods in Molecular Biology, vol 1255. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-2175-1_7

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  • DOI: https://doi.org/10.1007/978-1-4939-2175-1_7

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-2174-4

  • Online ISBN: 978-1-4939-2175-1

  • eBook Packages: Springer Protocols

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