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RNA-RNA Interactions between RNA Elements at the 5' end and at the Upstream of sgRNA of RNA Genome are Required for Potato virus X RNA Replication

  • Park, Mi-Ri (Department of Agricultural Biotechnology, Seoul National Nniversity) ;
  • Park, Sang-Ho (Department of Agricultural Biotechnology, Seoul National Nniversity) ;
  • Cho, Sang-Yun (Department of Agricultural Biotechnology, Seoul National Nniversity) ;
  • Hemenway, Cynthia L. (Department of Molecular and Structural Biochemistry, Box 7622, North Carolina State University) ;
  • Choi, Hong-Soo (Department of Plant Pathology, National Institute of Agricultural Science and Technology, RDA) ;
  • Sohn, Seong-Han (Cell and Genetics Division, National Institute of Agricultural Biotechnology, RDA) ;
  • Kim, Kook-Hyung (Department of Agricultural Biotechnology, Seoul National Nniversity)
  • Published : 2008.09.30

Abstract

RNA-RNA interactions and the dynamic RNA conformations are important regulators in virus replication in several RNA virus systems and may also involved in the regulation of many important virus life cycle phases, including translation, replication, assembly, and switches in these important stages. The 5' non-translated region of Potato virus X(PVX) contains multiple cis-acting elements that facilitate various viral processes. It has previously been proposed that RNA-RNA interactions between various RNA elements present in PVX RNA genome are required for PVX RNA accumulation(Hu et al., 2007; Kim and Hemenway, 1999). This model was based on the potential base-pairing between conserved sequence elements at the upstream of subgenomic RNAs(sgRNAs) and at the 5' and 3' end of RNA genome. We now provide more evidence that RNA-RNA base-pairing between elements present at the 5' end and upstream of each sgRNA is required for efficient replication of genomic and subgenomic plus-strand RNA accumulation. Site-directed mutations introduced at the 5' end of plus-strand RNA replication defective mutant(${\Delta}12$) increasing base-pairing possibility with conserved sequence elements located upstream of each sgRNAs restored genomic and subgenomic plus-strand RNA accumulation and caused symptom development in inoculated Nicotiana benthamiana plants. Serial passage of a deletion mutant(${\Delta}8$) caused more severe symptoms and restored wild type sequences and thus retained possible RNA-RNA base-pairing. Altogether, these results indicate that the RNA element located at the 5' end of PVX genome involved in RNA-RNA interactions and play a key role in high-level accumulation of plus-strand RNA in vivo.

Keywords

References

  1. Alvarez, D. E., Lodeiro, M. F., Luduena, S. J., Pietrasanta, L. I. and Gamarnik, A. V. 2005. Long-range RNA-RNA interactions circularize the dengue virus genome. J. Virol. 79:6631-6643 https://doi.org/10.1128/JVI.79.11.6631-6643.2005
  2. Hu, B., Pillai-Nair, N. and Hemenway, C. 2007. Long-distance RNA-RNA interactions between terminal elements and the same subset of internal elements on the potato virus X genome mediate minus- and plus-strand RNA synthesis. RNA 13:267-280 https://doi.org/10.1261/rna.243607
  3. Iwakawa, H. O., Kaido, M., Mise, K. and Okuno, T. 2007. cis-Acting core RNA elements required for negative-strand RNA synthesis and cap-independent translation are separated in the 3'-untranslated region of Red clover necrotic mosaic virus RNA1. Virology 369:168-181 https://doi.org/10.1016/j.virol.2007.07.016
  4. Karpova, O. V., Arkhipenko, M. V., Zaiakina, O. V., Nikitin, N. A., Kiseleva, O. I., Kozlovskii, S. V., Rodionova, N. P. and Atabekov, I. G. 2006. Translational regulation of potato virus X RNA-coat protein complexes: the key role of a coat protein N-terminal peptide. Mol. Biol. (Mosk) 40:703-710
  5. Kim, K.-H. and Hemenway, C. L. 1996. The 5' nontranslated region of potato virus X RNA affects both genomic and subgenomic RNA synthesis. J. Virol. 70:5533-5540
  6. Kim, K.-H. and Hemenway, C. L. 1997. Mutations that alter a conserved element upstream of the potato virus X triple block and coat protein genes affect subgenomic RNA accumulation. Virology 232:187-197 https://doi.org/10.1006/viro.1997.8565
  7. Kim, K.-H. and Hemenway, C. L. 1999. Long-distance RNARNA interactions and conserved sequence elements affect potato virus X plus-strand RNA accumulation. RNA 5:636-645 https://doi.org/10.1017/S1355838299982006
  8. Kim, K.-H., Kwon, S.-J. and Hemenway, C. L. 2002. Cellular protein binds to sequences near the 5' terminus of potato virus X RNA that are important for virus replication. Virology 301:305-312 https://doi.org/10.1006/viro.2002.1559
  9. Kunkel, T. A. 1985. Rapid and efficient site-specific mutagenesis without phenotypic selection. Proc. Natl. Acad. Sci. USA 82:488-492 https://doi.org/10.1073/pnas.82.2.488
  10. Kwon, S.-J., Park, M.-R., Kim, K.-W., Plante, C. A., Hemenway, C. L. and Kim, K.-H. 2005. cis-Acting sequences required for coat protein binding and in vitro assembly of Potato virus X. Virology 334:83-97 https://doi.org/10.1016/j.virol.2005.01.018
  11. Laemmli, U. K. 1970. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680-685 https://doi.org/10.1038/227680a0
  12. Lough, T. J., Lee, R. H., Emerson, S. J., Forster, R. L. and Lucas, W. J. 2006. Functional analysis of the 5' untranslated region of potexvirus RNA reveals a role in viral replication and cell-tocell movement. Virology 351:455-465 https://doi.org/10.1016/j.virol.2006.03.043
  13. Miller, E. D., Kim, K.-H. and Hemenway, C. L. 1999. Restoration of a stem-loop structure required for potato virus X RNA accumulation indicates selection for a mismatch and a GNRA tetraloop. Virology 260:342-353 https://doi.org/10.1006/viro.1999.9843
  14. Miller, E. D., Plante, C. A., Kim, K.-H., Brown, J. W. and Hemenway, C. L. 1998. Stem-loop structure in the 5' region of Potato virus X genome required for plus-strand RNA accumulation. J. Mol. Biol. 284:591-608 https://doi.org/10.1006/jmbi.1998.2174
  15. Mun, H.-Y., Park, M.-R., Lee, H.-B. and Kim, K.-H. 2008. Outbreak of Cucumber mosaic virus and Tomato spotted wilt virus on bell pepper grown in Jeonnam province in Korea. Plant Pathol. J. 24:93-96 https://doi.org/10.5423/PPJ.2008.24.1.093
  16. Olsthoorn, R. C., Mertens, S., Brederode, F. T. and Bol, J. F. 1999. A conformational switch at the 3' end of a plant virus RNA regulates viral replication. EMBO J. 18:4856-4864 https://doi.org/10.1093/emboj/18.17.4856
  17. Park, M.-R., Kwon, S.-J., Choi, H.-S., Hemenway, C. L. and Kim, K.-H. 2008. Mutations that alter a repeated ACCA element located at the 5' end of the Potato virus X genome affect RNA accumulation. Virology 378:133-141 https://doi.org/10.1016/j.virol.2008.05.004
  18. Park, S.-H. and Kim, K.-H. 2006. Agroinfiltration-based Potato virus X replications to dissect the requirements of viral infection. Plant Pathol. J. 22:386-390 https://doi.org/10.5423/PPJ.2006.22.4.386
  19. Ray, D., Wu, B. and White, K. A. 2003. A second functional RNA domain in the 5' UTR of the Tomato bushy stunt virus genome: intra- and interdomain interactions mediate viral RNA replication. RNA 9:1232-1245 https://doi.org/10.1261/rna.5630203
  20. Shen, R. and Miller, W. A. 2007. Structures required for poly(A) tail-independent translation overlap with, but are distinct from, cap-independent translation and RNA replication signals at the 3' end of Tobacco necrosis virus RNA. Virology 358:448-458 https://doi.org/10.1016/j.virol.2006.08.054
  21. Zhang, J., Zhang, G., Guo, R., Shapiro, B. A. and Simon, A. E. 2006. A pseudoknot in a preactive form of a viral RNA is part of a structural switch activating minus-strand synthesis. J. Virol. 80:9181-9191 https://doi.org/10.1128/JVI.00295-06

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  1. Nicotiana benthamiana protein, NbPCIP1, interacting with Potato virus X coat protein plays a role as susceptible factor for viral infection vol.386, pp.2, 2009, https://doi.org/10.1016/j.virol.2008.12.044