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Reduced function of the RNA-binding protein FPA rescues a T-DNA insertion mutant in the Arabidopsis ZHOUPI gene by promoting transcriptional read-through

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Abstract

T-DNA insertion mutants have been widely used to investigate plant gene functions. Unexpectedly, in several reported cases, the phenotype of T-DNA insertion mutations can be suppressed because of trans T-DNA interactions associated with epigenetic modification, which indicates that caution is needed when T-DNA mutants are used. In the present study, we characterized a novel process suppressing a T-DNA mutation. The spz2 (suppressor of zou 2) mutant was isolated as a suppressor of the phenotype of the zou-4 mutant caused by a T-DNA insertion in the first intron. The spz2 mutation partially recovered the native ZOU gene expression in the zou-4 background, but not in two other zou alleles, zou-2 and zou-3, with T-DNAs inserted in the exon and intron, respectively. The suppressed phenotype was inherited in a Mendelian fashion and is not associated with epigenetic modification. The recovery of the native ZOU gene expression in the spz2 zou-4 double mutant is caused by transcriptional read-through of the intronic T-DNA as a result of decreased proximal polyadenylation. SPZ2 encodes an RNA-binding protein, FPA, which is known to regulate polyadenylation site selection. This is the first example of FPA rescuing a T-DNA insertion mutation by affecting the polyadenylation site selection.

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Acknowledgments

This work was supported by the National Nature Science Foundation of China (grant nos. 31470286, 30970278 and 31271743) and the China National Transgenic Major Program (grant no. 2014ZX0800943B) and the Natural Science Foundation of Shandong Province of China (grant no. ZR2011CM020) and was also supported by the One Hundred Person Project of the Chinese Academy of Sciences.

Author contributions

S.X.Y., X.Z.F. and J.G. conceived the project and designed this work. Y.H.Z. and X.L. constructed the mutation library and performed map-based cloning. Y.H.Z., X.L. and C.X.W. performed transgenic, cell biological and other functional analyses. H.C.W. performed RNA sequencing analysis. S.X.Y., X.Z.F. and J.G. wrote this paper.

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    Authors and Affiliations

    1. Key Laboratory of Plant Stress Research, College of Life Science, Shandong Normal University, Jinan, 250014, People’s Republic of China

      Yaohua Zhang, Xin Li, Chunxia Wu & Xianzhong Feng

    2. Key Laboratory of Soybean Molecular Design Breeding, Northeast Institute of Geography and Agroecology, Chinese Academy of Sciences, Changchun, 130102, People’s Republic of China

      Yaohua Zhang, Xin Li, Haichao Wei, Suxin Yang & Xianzhong Feng

    3. Institute of Molecular Plant Sciences, University of Edinburgh, Daniel Rutherford Building, Max Born Crescent, Edinburgh, EH9 3BF, UK

      Justin Goodrich

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    1. Yaohua Zhang
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    Yaohua Zhang and Xin Li contributed equally to this work.

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    Zhang, Y., Li, X., Goodrich, J. et al. Reduced function of the RNA-binding protein FPA rescues a T-DNA insertion mutant in the Arabidopsis ZHOUPI gene by promoting transcriptional read-through. Plant Mol Biol 91, 549–561 (2016). https://doi.org/10.1007/s11103-016-0487-2

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