Transgenerational Transmission of Radiation-Induced Expression Patterns of Arabidopsis Thaliana (L.) Heynh. Rad51 and Rad1 Genes

Öz

Transcription rates of the genes AtKu70, AtRAD51, AtRad1, involved in maintaining Arabidopsis thaliana genome stability, in relation to the modification of phenotypic characteristics in irradiated plants and their progeny after the action of acute and fractionated X-ray radiation were studied. Differences in the transcription rate were measured by densitometric analysis of cDNA, synthesized by reverse transcription at the template of mRNAs, extracted from fresh leaves after 2 hours irradiation treatment. The doses 3 Gy, 12 Gy, 15 Gy and 21 Gy with 1.48 Gy/s specific dose rate were applied. Significant correlation between phenotype modifications in F0 and F1 generations, between phenotype traits and caretaker genes activity in irradiated F0 plants were shown. Also preservation of changes in the pattern of AtRad1 and AtRAD51 but not AtKu70 expression in F1 plant leaves had been revealed. Changes in F1 compared with F0 generation do not correspond to the extrapolation of dependence between the phenotypic modifications and DNA repair genes transcription rate in the leaves of irradiated plants. Based on the obtained data it could be suggested that the altered transcriptional activity of AtRAD51 and AtRad1 reflects the transfer of DNA lesions from parent to offspring.

Anahtar Kelimeler

• Genes expression
• X-ray irradiation
• DNA repair
• Transgenerational inheritance

Transgenerational Transmission of Radiation-Induced Expression Patterns of Arabidopsis Thaliana (L.) Heynh. Rad51 and Rad1 Genes

Abstract

Transcription rates of the genes AtKu70, AtRAD51, AtRad1, involved in maintaining Arabidopsis thaliana genome stability, in relation to the modification of phenotypic characteristics in irradiated plants and their progeny after the action of acute and fractionated X-ray radiation were studied. Differences in the transcription rate were measured by densitometric analysis of cDNA, synthesized by reverse transcription at the template of mRNAs, extracted from fresh leaves after 2 hours irradiation treatment. The doses 3 Gy, 12 Gy, 15 Gy and 21 Gy with 1.48 Gy/s specific dose rate were applied. Significant correlation between phenotype modifications in F₀ and F₁ generations, between phenotype traits and caretaker genes activity in irradiated F₀ plants were shown. Also preservation of changes in the pattern of AtRad1 and AtRAD51 but not AtKu70 expression in F₁ plant leaves had been revealed. Changes in F₁ compared with F₀ generation do not correspond to the extrapolation of dependence between the phenotypic modifications and DNA repair genes transcription rate in the leaves of irradiated plants. Based on the obtained data it could be suggested that the altered transcriptional activity of AtRAD51 and AtRad1 reflects the transfer of DNA lesions from parent to offspring.

Keywords

• Genes expression
• X-ray irradiation
• DNA repair
• Transgenerational inheritance

References

1. Mannuss, A., Trapp, O., Puchta, H. (2012). Gene Regulation in Response to DNA Damage, Biochimica et Biophysica Acta, 1819(2), 154-165. http://dx.doi.org/10.1016/j.bbagrm.2011.08.003
2. Vannier, J.-B., Depeiges, A., White, C., Gallego, M.E. (2009). ERCC1/XPF Protects Short Telomeres from Homologous Recombination in Arabidopsis thaliana, PLOS Genetics, 5(2), e1000380. http://dx.doi.org/10.1371/journal.pgen.1000380
3. Yoshiyama, K., Sakaguchi, K., Kimura, S. (2013). DNA Damage Response in Plants: Conserved and Variable Response Compared to Animals, Biology, 2 (4), 1338-1356. http://dx.doi.org/10.3390/biology2041338
4. Boulton, S., Jackson, S. (1998) Components of the Ku-dependent non-homologous end-joining pathway are involved in telomeric length maintenance and telomeric silencing, The EMBO Journal, 17(6), 1819-1828. http://dx.doi.org/10.1093/emboj/17.6.1819
5. Frankenberg-Schwager, M., Gebauer, A., Koppe, C., Wolf, H., Pralle, E., Frankenberg, D. (2009). Single-strand annealing, conservative homologous recombination, nonhomologous DNA end joining, and the cell cycle-dependent repair of DNA double-strand breaks induced by sparsely or densely ionizing radiation, Radiation Research, 171(3), 265-273. http://dx.doi.org/10.1667/RR0784.1
6. Lieber, M.R. (2010). NHEJ and Its Backup Pathways: Relation to Chromosomal Translocations, Nature Structural & Molecular Biology, 17(4), 393-395. http://dx.doi.org/10.1038/nsmb0410-393
7. Waterworth, W., Drury, G., Bray, C., West, C. (2011). Repairing Breaks in the Plant Genome: The Importance of Keeping It Together, New Phytologist, 192 (4), 805-822. http://dx.doi.org/10.1111/j.1469-8137.2011.03926.x
8. Li, J., Harper, L.C., Golubovskaya, I., Wang, C.R., Weber, D., Meeley, R.B., McElver, J., Bowen, B., Cande, W.Z., Schnable, P.S. (2007). Functional Analysis of Maize RAD51 in Meiosis and Double–Strand Break Repair, Genetics, 176(3), 1469-1482. http://dx.doi.org/10.1534/genetics.106.062604

9. Ma, W., Westmoreland J. W., Resnick M. A. (2013). Homologous recombination rescues ssDNA gaps generated by nucleotide excision repair and reduced translesion DNA synthesis in yeast G2 cells, PNAS, 110(31), 2895-2904. http://dx.doi.org/10.1073/pnas.1301676110
10. Besse, B., Olaussen, K.A., Soria, J.C. (2013). ERCC1 and RRM1: ready for prime time?, Journal of Clinical Oncology, 31(8), 1050-1060. http://dx.doi.org/10.1200/JCO.2012.43.0900
11. Hwang, J.-Y., Smith, S., & Myung, K. (2005). The Rad1-Rad10 Complex Promotes the Production of Gross Chromosomal Rearrangements from Spontaneous DNA Damage in Saccharomyces cerevisiae, Genetics, 169(4), 1927-1937. http://dx.doi.org/10.1534/genetics.104.039768
12. Ma, J.L., Kim, E.M., Haber, J.E., Lee, S.E. (2003). Yeast Mre11 and Rad1 proteins define a Ku-independent mechanism to repair double-strand breaks lacking overlapping end sequences, Molecular and Cellular Biology, 23(23), 8820-8828. http://dx.doi.org/10.1128/MCB.23.23.8820-8828.2003
13. McVey, M., Lee, S.E. (2008). MMEJ repair of double-strand breaks (director's cut): deleted sequences and alternative endings, Trends in Genetics, 24(11), 529-538. http://dx.doi.org/10.1016/j.tig.2008.08.007
14. Scuric, Z., Chan, C.Y., Hafer, K., Schiestl, R.H. (2009). Ionizing Radiation Induces Microhomology-Mediated End Joining in trans in Yeast and Mammalian Cells, Radiation Research, 171(4), 454-463. http://dx.doi.org/10.1667/RR1329.1
15. Zhang, Y., Rohde, L.H., Wu, H. (2009). Involvement of nucleotide excision and mismatch repair mechanisms in double strand break repair, Current Genomics, 10(4), 250-258. http://dx.doi.org/10.2174/138920209788488544
16. Litvinov, S., Rashydov, N. (2017) The transcriptional response of Arabidopsis thaliana L. AtKu70, AtRAD51 and AtRad1 genes to X-rays, Journal of Agricultural Science and Technology A, 7 (1), 52-60. http://dx.doi.org/10.17265/2161-6256/2017.01.008
17. Bradford, W., Cahoon, L., Freel, S., Mays Hoopes, L., Eckdahl, T. (2005) An Inexpensive Gel Electrophoresis-Based Polymerase Chain Reaction Method for Quantifying mRNA Levels, Cell Biology Education, 4(2), 157-168. http://dx.doi.org/10.1187/cbe.04-09-0051
18. Litvinov, S. (2014). Effects of Chronic Exposure of Seeds and Seedlings of Arabidopsis thaliana by Low Doses of γ-Radiation on Plant Growth and Development, Nuclear Physics and Atomic Energy, 15(4), 406-414.