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Identification of a plant-specific Zn2+-sensitive ribonuclease activity

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Abstract

Ribonucleases (RNases) play a variety of cellular and biological roles in all three domains of life. In an attempt to perform RNA immuno-precipitation assays of Arabidopsis proteins, we found an EDTA-dependent RNase activity from Arabidopsis suspension tissue cultures. Further investigations proved that the EDTA-dependent RNase activity was plant specific. Characterization of the RNase activity indicated that it was insensitive to low pH and high concentration of NaCl. In the process of isolating the activity with cation exchange chromatography, we found that the EDTA dependency of the activity was lost. This led us to speculate that some metal ions, which inhibited the RNase activity, may be removed during cation exchange chromatography so that the nuclease activity was released. The EDTA dependency of the activity could be due to the ability of the EDTA chelating those metal ions, mimicking the effect of the cation exchange chromatography. Indeed, Zn2+ strongly inhibited the activity, and the inhibition could be released by EDTA based on both in-solution and in-gel assays. In-gel assays identified two RNase activity bands. Mass spectrometry assays of those activity bands revealed more than 20 proteins. However, none of them has an apparent known nuclease domain, suggesting that one or more of those proteins might possess a currently uncharacterized nuclease domain. Our results may shed light on RNA metabolism in plants by introducing a novel plant-specific RNase activity.

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Acknowledgments

We highly appreciate constructive suggestions of anonymous reviewers in terms of potential identity of the RNase activity. This work was supported in part by a grant from US National Institute of Health (1R15GM07719201A1 to QQL), and by a grant from Ohio Plant Biotech Consortium (to QQL and DX).

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Authors and Affiliations

  1. Department of Botany, Miami University, Oxford, OH, 45056, USA

    Denghui Xing & Qingshun Quinn Li

  2. Department of Chemistry and Biochemistry, Miami University, Oxford, OH, 45056, USA

    Shuisong Ni & Michael A. Kennedy

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  1. Denghui Xing
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  2. Shuisong Ni
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  3. Michael A. Kennedy
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  4. Qingshun Quinn Li
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Correspondence to Qingshun Quinn Li.

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Xing, D., Ni, S., Kennedy, M.A. et al. Identification of a plant-specific Zn2+-sensitive ribonuclease activity. Planta 230, 819–825 (2009). https://doi.org/10.1007/s00425-009-0986-3

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