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Expression of a Tardigrade Dsup Gene Enhances Genome Protection in Plants

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Abstract

DNA damage is one of the most impactful events in living organisms, leading to DNA sequence changes (mutation) and disruption of biological processes. A study has identified a protein called Damage Suppressor Protein (Dsup) in the tardigrade Ramazzotius varieornatus that has shown to reduce the effects of radiation damage in human cell cultures (Hashimoto in Nature Communications 7:12808, 2016). We have generated tobacco plants that express the codon-optimized tardigrade Dsup gene and examined their responses when treated with mutagenic chemicals, ultraviolet (UV) and ionizing radiations. Our studies showed that compared to the control plants, the Dsup-expressing plants grew better in the medium containing mutagenic ethylmethane sulfonate (EMS). RT-qPCR detected distinct expression patterns of endogenous genes involved in DNA damage response and repair in the Dsup plants in response to EMS, bleomycin, UV-C and X-ray radiations. Comet assays revealed that the nuclei from the Dsup plants appeared more protected from UV and X-ray damages than the control plants. Overall, our studies demonstrated that Dsup gene expression enhanced tolerance of plants to genomutagenic stress. We suggest the feasibility of exploring genetic resources from extremotolerant species such as tardigrades to impart plants with tolerance to stressful environments for future climate changes and human space endeavors.

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References

  1. Oztas, O., Selby, C. P., Sancar, A., & Adebali, O. (2018). Genome-wide excision repair in Arabidopsis is coupled to transcription and reflects circadian gene expression patterns. Nature Communications. https://doi.org/10.1038/s41467-018-03922-5.

    Article  PubMed  PubMed Central  Google Scholar 

  2. Britt, A. B. (1996). DNA damage and repair in plants. Annual Review of Plant Physiology and Plant Molecular Biology, 47, 75–100. https://doi.org/10.1146/annurev.arplant.47.1.75.

    Article  PubMed  CAS  Google Scholar 

  3. Hu, Z., Cools, T., & De Veylder, L. (2016). Mechanisms used by plants to cope with DNA damage. Annual Review of Plant Biology, 67, 439–462. https://doi.org/10.1146/annurev-arplant-043015-111902.

    Article  PubMed  CAS  Google Scholar 

  4. Marcu, D., Damian, G., Cosma, C., & Cristea, V. (2013). Gamma radiation effects on seed germination, growth and pigment content, and ESR study of induced free radicals in maize (Zea mays). Journal of Biological Physics, 39, 625–634. https://doi.org/10.1007/s10867-013-9322-z.

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  5. Sharma, P., Jha, A. B., Dubey, R. S., & Pessarakli, M. (2012). Reactive oxygen species, oxidative damage, and antioxidative defense mechanism in plants under stressful conditions. Journal of Botany, 2012, 1–26. https://doi.org/10.1155/2012/217037.

    Article  CAS  Google Scholar 

  6. Gupta, D. K., Palma, J. M., & Corpas, F. J. (2015). Reactive oxygen species and oxidative damage in plants under stress. Heidelberg: Springer. https://doi.org/10.1007/978-3-319-20421-5.

    Book  Google Scholar 

  7. Khursheed, S., Raina, A., Laskar, R. A., & Khan, S. (2018). Effect of gamma radiation and EMS on mutation rate: their effectiveness and efficiency in faba bean (Vicia faba L.). Caryologia, 71, 397–404. https://doi.org/10.1080/00087114.2018.1485430.

    Article  Google Scholar 

  8. Rošić, S., Amouroux, R., Requena, C. E., Gomes, A., Emperle, M., Beltran, T., et al. (2018). Evolutionary analysis indicates that DNA alkylation damage is a byproduct of cytosine DNA methyltransferase activity. Nature Genetics, 50, 452–459. https://doi.org/10.1038/s41588-018-0061-8.

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  9. Drabløs, F., Feyzi, E., Aas, P. A., Vaagbø, C. B., Kavli, B., Bratlie, M. S., et al. (2004). Alkylation damage in DNA and RNA—Repair mechanisms and medical significance. DNA Repair, 3, 1389–1407. https://doi.org/10.1016/j.dnarep.2004.05.004.

    Article  PubMed  CAS  Google Scholar 

  10. Steighner, R. J., & Povirk, L. F. (1990). Bleomycin-induced DNA lesions at mutational hot spots: Implications for the mechanism of double-strand cleavage. Proceedings of the National Academy of Sciences of the United States of America, 87, 8350–8354. https://doi.org/10.1073/pnas.87.21.8350.

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  11. Chen, J., & Stubbe, J. A. (2005). Bleomycins: Towards better therapeutics. Nature Reviews Cancer, 5, 102–112. https://doi.org/10.1038/nrc1547.

    Article  PubMed  CAS  Google Scholar 

  12. Bolzán, A. D., & Bianchi, M. S. (2018). DNA and chromosome damage induced by bleomycin in mammalian cells: An update. Mutation Research—Reviews in Mutation Research, 775, 51–62. https://doi.org/10.1016/j.mrrev.2018.02.003.

    Article  PubMed  CAS  Google Scholar 

  13. Horikawa, D. D., Kunieda, T., Abe, W., Watanabe, M., Nakahara, Y., Yukuhiro, F., et al. (2008). Establishment of a rearing system of the extremotolerant tardigrade Ramazzottius varieornatus: A new model animal for astrobiology. Astrobiology, 8, 549–556. https://doi.org/10.1089/ast.2007.0139.

    Article  PubMed  CAS  Google Scholar 

  14. Guidetti, R., Rizzo, A. M., Altiero, T., & Rebecchi, L. (2012). What can we learn from the toughest animals of the Earth? Water bears (tardigrades) as multicellular model organisms in order to perform scientific preparations for lunar exploration. Planetary and Space Science, 74, 97–102. https://doi.org/10.1016/j.pss.2012.05.021.

    Article  Google Scholar 

  15. Hashimoto, T., & Kunieda, T. (2017). DNA protection protein, a novel mechanism of radiation tolerance: Lessons from tardigrades. Life, 7, 1–11. https://doi.org/10.3390/life7020026.

    Article  CAS  Google Scholar 

  16. Hashimoto, T., Horikawa, D. D., Saito, Y., Kuwahara, H., Kozuka-Hata, H., Shin-I, T., et al. (2016). Extremotolerant tardigrade genome and improved radiotolerance of human cultured cells by tardigrade-unique protein. Nature Communications, 7, 12808. https://doi.org/10.1038/ncomms12808.

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  17. Kirke, J., Kaplan, N., Velez, S., Jin, X. L., Vichyavichien, P., & Zhang, X. H. (2018). Tissue-preferential activity and induction of the pepper capsaicin synthase PUN1 promoter by wounding, heat and metabolic pathway precursor in tobacco and tomato plants. Molecular Biotechnology, 60, 194–202. https://doi.org/10.1007/s12033-018-0060-0.

    Article  PubMed  CAS  Google Scholar 

  18. Murashige, T., & Skoog, F. (1962). A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiologia Plantarum, 15, 473–497.

    Article  CAS  Google Scholar 

  19. Zhang, X.-H., Takagi, H., & Widholm, J. M. (2004). Expression of a novel yeast gene that detoxifies the proline analog azetidine-2-carboxylate confers resistance during tobacco seed germination, callus and shoot formation. Plant Cell Reports, 22, 615–622.

    Article  CAS  Google Scholar 

  20. Tsai, F.-Y., Zhang, X.-H., Ulanov, A., & Widholm, J. M. (2010). The application of the yeast N-acetyltransferase MPR1 gene and the proline analogue L-azetidine-2-carboxylic acid as a selectable marker system for plant transformation. Journal of Experimental Botany, 61, 2561–2573.

    Article  CAS  Google Scholar 

  21. Hill, W., Jin, X.-L., & Zhang, X.-H. (2016). Expression of an arctic chickweed dehydrin, CarDHN, enhances tolerance to abiotic stress in tobacco plants. Plant Growth Regulation, 80, 323–334.

    Article  CAS  Google Scholar 

  22. Schmidt, G. W., & Delaney, S. K. (2010). Stable internal reference genes for normalization of real-time RT-PCR in tobacco (Nicotiana tabacum) during development and abiotic stress. Molecular Genetics and Genomics, 283, 233–241.

    Article  CAS  Google Scholar 

  23. Zhang, X.-H., Keating, P., Wang, X.-W., Huang, Y.-H., Martin, J., Hartmann, J. X., et al. (2014). Production of functional native human interleukin-2 in tobacco chloroplasts. Molecular Biotechnology, 56, 369–379.

    Article  CAS  Google Scholar 

  24. Zhang, X.-H., Webb, J., Huang, Y.-H., Lin, L., Tang, R.-S., & Liu, A. (2011). Hybrid Rubisco of tomato large subunits and tobacco small subunits is functional in tobacco plants. Plant Science, 180, 480–488.

    Article  CAS  Google Scholar 

  25. Sikorskaite, S., Rajamäki, M.-L., Baniulis, D., Stanys, V., & Valkonen, J. P. T. (2013). Protocol: Optimised methodology for isolation of nuclei from leaves of species in the Solanaceae and Rosaceae families. Plant Methods, 9, 31.

    Article  CAS  Google Scholar 

  26. Olive, P. L., & Banáth, J. P. (2006). The comet assay: A method to measure DNA damage in individual cells. Nature Protocols, 1, 23–29. https://doi.org/10.1038/nprot.2006.5.

    Article  PubMed  CAS  Google Scholar 

  27. Georgieva, M., & Stoilov, L. (2008). Assessment of DNA strand breaks induced by bleomycin in barley by the comet assay. Environmental and Molecular Mutagenesis, 49, 381–387. https://doi.org/10.1002/em.20396.

    Article  PubMed  CAS  Google Scholar 

  28. Hille, J., Verheggen, F., Roelvink, P., Franssen, H., van Kammen, A., & Zabel, P. (1986). Bleomycin resistance: A new dominant selectable marker for plant cell transformation. Plant Molecular Biology, 7, 171–176.

    Article  CAS  Google Scholar 

  29. Kumaravel, T. S., Vilhar, B., Faux, S. P., & Jha, A. N. (2009). Comet assay measurements: A perspective. Cell Biology and Toxicology, 25, 53–64. https://doi.org/10.1007/s10565-007-9043-9.

    Article  PubMed  CAS  Google Scholar 

  30. Yektaeian, N., Mehrabani, D., Sepaskhah, M., Zare, S., Jamhiri, I., & Hatam, G. (2019). Lipophilic tracer Dil and fluorescence labeling of acridine orange used for Leishmania major tracing in the fibroblast cells. Heliyon, 5, e03073. https://doi.org/10.1016/j.heliyon.2019.e03073.

    Article  PubMed  PubMed Central  Google Scholar 

  31. Manova, V., & Gruszka, D. (2015). DNA damage and repair in plants—From models to crops. Frontiers in Plant Science, 6, 1–26. https://doi.org/10.3389/fpls.2015.00885.

    Article  Google Scholar 

  32. Fulcher, N., & Sablowski, R. (2009). Hypersensitivity to DNA damage in plant stem cell niches. Proceedings of the National Academy of Sciences of the United States of America, 106, 20984–20988. https://doi.org/10.1073/pnas.0909218106.

    Article  PubMed  PubMed Central  Google Scholar 

  33. Collins, A. R. (2004). The comet assay for DNA damage and repair: Principles, applications, and limitations. Applied Biochemistry and Biotechnology—Part B Molecular Biotechnology, 26, 249–261. https://doi.org/10.1385/MB:26:3:249.

    Article  CAS  Google Scholar 

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Acknowledgements

The work was supported in part by the Undergraduate Research Grants from Florida Atlantic University (FAU) and Miami Dade College School of Science STEM Summer Research Program. Undergraduate students Ronscardy Mondesir, Andrew Adeyiga, Tahoe Albergo, Andrew Balsamo, Nicholas Nifakos, Milove Jeannot, Amanda Lam, Nicholas Pizzo and Mohamed Abutineh participated in various stages of experiments of this project. We thank Sophia Zheng for initial discussion of using Dsup as a possible high school science project. We also thank Drs. Patricia Keating and James Hartmann for exploring with flow cytometry assays, A.D. Henderson University High School, FAU Owls Imaging Lab for CT scanning and FAU School of Medicine imaging center for fluorescence microscopy.

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  1. Department of Biological Sciences, Florida Atlantic University, Boca Raton, FL, 33431, USA

    Justin Kirke, Xiao-Lu Jin & Xing-Hai Zhang

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  1. Justin Kirke
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XHZ conceived and designed the experiments. JK, XLJ, and XHZ performed the experiments. XHZ and JK wrote the manuscript with comments from all co-authors.

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Correspondence to Xing-Hai Zhang.

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Kirke, J., Jin, XL. & Zhang, XH. Expression of a Tardigrade Dsup Gene Enhances Genome Protection in Plants. Mol Biotechnol 62, 563–571 (2020). https://doi.org/10.1007/s12033-020-00273-9

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