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Investigation of Deletion Variation and Methylation Patterns in the 5' LTR of Porcine Endogenous Retroviruses

  • Jung, K.C. (Division of Animal Science and Resources, Research Center for Transgenic Cloned Pigs Chungnam National University) ;
  • Simond, D.M. (Centre for Transplantation and Renal Research, Westmead Millennium Institute) ;
  • Moran, C. (Faculty of Veterinary Science, University of Sydney) ;
  • Hawthorne, W.J. (Centre for Transplantation and Renal Research, Westmead Millennium Institute) ;
  • Jeon, J.T. (Division of Applied Life Science, Gyeongsang National University) ;
  • Jin, D.I. (Division of Animal Science and Resources, Research Center for Transgenic Cloned Pigs Chungnam National University) ;
  • Lee, J.H. (Division of Animal Science and Resources, Research Center for Transgenic Cloned Pigs Chungnam National University)
  • Received : 2008.01.25
  • Accepted : 2008.07.09
  • Published : 2008.11.01

Abstract

The xenotransplantation of pig organs and cells can be related with a risk of transmission of infectious diseases to human. Previous findings indicate that the regulatory region of PERV for retroviral transcription, replication and integration into the cellular DNA is located on the 5' Long Terminal Repeat (LTR). The objective of this study is the investigation of methylation and deletion status of the PERV 5' LTR region which can be used for regulating PERV expression. We compared the sequences of genomic DNA and bisulfite-treated genomic DNA from PK-15 cells expressing PERV to observe the methylation status of the 5' LTR. Our results showed that the CpG sites of U3 were methylated and methylation was inconsistent in the R and U5 regions. Also, variable numbers of 18 bp repeats and 21 bp repeats were detected on 5' LTR by sequencing analysis. The consistent U3 methylation might be indicative of host suppression of expression of the retroviruses.

Keywords

Pigs;Long Terminal Repeat (LTR);Porcine Endogenous Retroviruses (PERVs);Xenotransplantation

Acknowledgement

Supported by : Rural Development Administration

References

  1. Yoder, J. A., C. P. Walsh and T. H. Bestor. 1997. Cytosine methylation and the ecology of intragenomic parasites. Trends Genet. 13:335-340. https://doi.org/10.1016/S0168-9525(97)01181-5
  2. Dieckhoff, B., A. Karlas, A. Hofmann, W. A. Kues, B. Petersen, A. Pfeifer, H. Niemann, R. Kurth and J. Denner. 2007. Inhibition of porcine endogenous retroviruses (PERVs) in primary porcine cells by RNA interference using lentiviral vectors. Arch. Virol. 152:629-634. https://doi.org/10.1007/s00705-006-0868-y
  3. Hanecak, R., S. Mittal, B. R. Davis and H. Fan. 1986. Generation of infectious Moloney murine leukemia viruses with deletions in the U3 portion of the long terminal repeat. Mol. Cell. Biol. 6:4634-4640. https://doi.org/10.1128/MCB.6.12.4634
  4. Lavie, L., M. Kitova, E. Maldener, E. Meese and J. Mayer. 2005. CpG methylation directly regulates transcriptional activity of the human endogenous retrovirus family HERV-K (HML-2). J. Virol. 79:876-883. https://doi.org/10.1128/JVI.79.2.876-883.2005
  5. Le Tissier, P., J. P. Stoye, Y. Takeuchi, C. Patience and R. A. Weiss. 1997. Two sets of human-tropic pig retrovirus. Nature 16:681-682.
  6. Scheef, G., N. Fischer, U. Krach and R. R. Tonjes. 2001. The number of a U3 repeat box acting as an enhancer in long terminal repeats of polytropic replication-competent porcine endogenous retroviruses dynamically fluctuates during serial virus passages in human cells. J. Virol. 75:6933-6940. https://doi.org/10.1128/JVI.75.15.6933-6940.2001
  7. Wilson, C. A., S. Laeeq, A. Ritzhaupt, W. Colon-Moran and F. K. Yoshimura. 2003. Sequence analysis of porcine endogenous retrovirus long terminal repeats and identification of transcriptional regulatory regions. J. Virol. 77:142-149. https://doi.org/10.1128/JVI.77.1.142-149.2003
  8. Bednarik, D. P., J. A. Cook and P. M. Pitha. 1990. Inactivation of the HIV LTR by DNA CpG methylation: evidence for a role in latency. EMBO J. 9:1157-1164.
  9. Bednarik, D. P., J. D. Mosca and N. B. Raj. 1987. Methylation as a modulator of expression of human immunodeficiency virus. J. Virol. 61:1253-1257.
  10. Bird, A. P. and A. P. Wolffe. 1999. Methylation-induced repression-belts, braces, and chromatin. Cell 99:451-454. https://doi.org/10.1016/S0092-8674(00)81532-9
  11. Cheung, P., C. D. Allis and P. Sassone-Corsi. 2000. Signaling to chromatin through histone modifications. Cell 103:263-271. https://doi.org/10.1016/S0092-8674(00)00118-5
  12. Jung, K. C., S. L. Yu, T. H. Kim, J. T. Jeon, C. Rogel-Gaillard, C. S. Park, D. I. Jin, C. Moran and J. H. Lee. 2007. Insertional variations of two porcine endogenous retroviruses (PERVs) in Korean native pigs and asian wild boars. Asian-Aust. J. Anim. Sci. 20:461-465. https://doi.org/10.5713/ajas.2007.461
  13. Wilson, C. A., S. Wong, J. Muller, C. E. Davidson, T. M. Rose and P. Burd. 1998. Type C retrovirus released from porcine primary peripheral blood mononuclear cells infects human cells. J. Virol. 72:3082-3087.

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