Journal of Life Science (생명과학회지)

Korean Society of Life Science (한국생명과학회)
권호 표지
DOI QR코드

DOI QR Code

Biosenesis of Epstein-Barr Virus MicroRNAs in B Cells

B 세포에서 Epstein-Barr virus microRNA들의 전사 및 성숙

  • Kim Do Nyun (Research Institute of Immunobiology, Department of Biomedical Sciences, Catholic University of Korea) ;
  • Oh Sang Taek (Research Institute of Immunobiology, Department of Biomedical Sciences, Catholic University of Korea) ;
  • Lee Jae Myun (Department of Microbiology, Yonsei University) ;
  • Lee Won-Keun (Division of Bioscience and Bioinformatics, Myongji University) ;
  • Lee Suk Kyeong (Research Institute of Immunobiology, Department of Biomedical Sciences, Catholic University of Korea)
  • 김도년 (가톨릭대학교 의과대학 생명의과학과 면역생물학연구소) ;
  • 오상택 (가톨릭대학교 의과대학 생명의과학과 면역생물학연구소) ;
  • 이재면 (연세대학교 의과대학 미생물하교실) ;
  • 이원근 (명지대학교 생명과학정보학부) ;
  • 이숙경 (가톨릭대학교 의과대학 생명의과학과 면역생물학연구소)
  • Published : 2005.12.01
Download PDF
⟨ Previous Next ⟩

Abstract

We investigated microRNA (miRNA) biogenesis of Epstein-Barr virus (EBV) which is the first virus shown to produce viral miRNAs. As expected, expression of all the reported EBV miRNAs were detected by Northen blot in an EBV-infected B cell line, B95-8; BHRF1-1, BHIU1-2, BHRF1-3, BART1, and BART2. The putative EBV pri-miRWAs and pre-miRNAs predicted from the known mature EBV miRNA sequences were detected by RT-PCR in B95-8 cells. Many animal miRNA genes exist as clusters of 2-7 genes and they are expressed polycistronically. As the EBV miRNAs are clustered in two regions of the EBV genome, we examined whether these clustered EBV miRNA genes are also expressed polycistronically. A long polycistronic transcript with the expected size (1602 bp) corresponding to the BHRF1-1~BHRF1-2~BHRF1-3 was amplified. However, any polycistronic transcript containing both BART1 and BART2 was detectable in B95-8. These results suggest that EBV miRNAs may be processed in a similar way with animal miRNAs and that some of the clustered EBV miRNAs can be transcribed polycistronically.

우리는 바이러스로는 최초로 miRNA를 생성한다고 보고된 Epstein-Barr virus (EBV)를 대상으로 EBV miRNA biogenesis를 연구했다. 먼저 EVB에 감염된 B 세포인 B95-8로부터 보고된 5 종의 EBV miRNA들인 BHRF1-1, BHRF1-2, BHRF1-3, BART1, 및 BART2 모두가 발현됨을 확인하였다. 그 다음 성숙된 EBV miRNA 서열로부터 예측되는 pri- 와 pre-miRNA들이 B95-8에서 각각 검출되어 EBV miRNA들도 이미 알려진 동물세포 miRNA와 유사한 생성 및 성숙과정을 거칠 가능성을 확인하였다 게놈 상에 2~7개씩 밀집하여 존재하는 동물세포 miRNA들과 유사하게 EBV 게놈의 2부 위에 밀집해서 존재하는 EBV miRNA들도 polycistronic하게 발현되는지 조사한 결과 B95-8에서 BHRF1-1, BHRF1-2 및 BHRF1-3를 포함하는 1,602 뉴클레오타이드의 긴 전사체가 RT-PCR로 확인되었다. 반면 BART1과 BART2는 mono cistronic하게 전사될 가능성이 확인되었다. 본 연구를 통하여 EBV miRNA들도 동물세포 miRNA들과 유사하게 poly cistronic 하게 발현될 수 있으며 pri-와 pre-miRNA 과정을 거쳐 성숙된 miRNA로 생성될 가능성을 확인하였다.

Keywords

References

  1. Aman, P., B. Ehlin-Henriksson and G. Klein, 1984, Epstein-Barr virus susceptibility of normal human B lymphocyte populations. J. Exp. Med. 159, 208-220 https://doi.org/10.1084/jem.159.1.208
  2. Ambros, V., B. Bartel, D. P. Bartel, C. B. Burge, J. C. Carrington, X. Chen, G. Dreyfuss, S. R. Eddy, S. GriffithsJones, M. Marshall, M. Matzke, G. Ruvkun and T. Tuschl. 2003. A uniform system for microRNA annotation. RNA 9, 277-279 https://doi.org/10.1261/rna.2183803
  3. Bartel, D. P. 2004. MicroRNAs: genomics, biogenesis, mechanism, and function. Cell 116, 281-297 https://doi.org/10.1016/S0092-8674(04)00045-5
  4. Bernstein, E., A. A. Caudy, S. M. Hammond and G. J. Hannon. 2001. Role for a bidentate ribonuclease in the initiation step of RNA interference. Nature 409, 363-366 https://doi.org/10.1038/35053110
  5. Bohnsack, M. T., K. Czaplinski and D. Gorlich. 2004. Exportin 5 is a RanGTP-dependent dsRNA-binding protein that mediates nuclear export of pre-miRNAs. RNA 10, 185-191 https://doi.org/10.1261/rna.5167604
  6. Chen, H., J. Huang, F. Y. we. G. Liao, L. Hutt-Fletcher and S. D. Hayward. 2005. Regulation of expression of the Epstein-Barr virus BamHI-A rightward transcripts. J. Virol. 79, 1724-1733 https://doi.org/10.1128/JVI.79.3.1724-1733.2005
  7. Cullen, B. R. 2004. Transcription and processing of human microRNA precursors. Mol. Cell 16, 861-865 https://doi.org/10.1016/j.molcel.2004.12.002
  8. de Jesus, O., P. R. Smith, L. C. Spender, C. Elgueta Karstegl, H. H. Niller, D. Huang and P. J. Farrell. 2003. Updated Epstein-Barr virus (EBV) DNA sequence and analysis of a promoter for the BART (CST, BARFO) RNAs of EBV. J. Gen. Virol. 84, 1443-1450 https://doi.org/10.1099/vir.0.19054-0
  9. Denli, A. M., B. B. Tops, R. H. Plasterk, R. F. Ketting and G. J. Hannon. 2004. Processing of primary microRNAs by the Microprocessor complex. Nature 432, 231-235 https://doi.org/10.1038/nature03049
  10. Gregory, R. I., K. P. Yan, G. Amuthan, T. Chendrimada, B. Doratotai, N. Cooch and R. Shiekhattar. 2004. The Microprocessor complex mediates the genesis of microRNAs. Nature 432,235-240 https://doi.org/10.1038/nature03120
  11. Grishok, A., A. E. Pasquinelli; D.Conte, N. Li. S. Parrish, I. Ha, D. L. Baillie, A. Fire, G. Ruvkun and C. C. Mello. 2001. Genes and mechanisms related to RNA interference regulate expression of the small temporal RNAs that control C. elegans developmental timing. Cell 106, 23-34 https://doi.org/10.1016/S0092-8674(01)00431-7
  12. Han, J., Y. Lee, K. H. Yeom, Y. K. Kim, H.Jin and V. N. Kim. 2004. The Drosha-DGCR8 complex in primary microRNA processing. Genes Dev. 18, 3016-3027 https://doi.org/10.1101/gad.1262504
  13. Hayes, D. P., A. A. Brink, M. B. Vervoort, J. M. Middeldorp, C. J. Meijer and A. J. van den Brule. 1999. Expression of Epstein-Barr virus (EBV) transcripts encoding homologues to important human proteins in diverse EBV associated diseases. Molecular Pathology 52, 97-103 https://doi.org/10.1136/mp.52.2.97
  14. He, L., J. M. Thomson, M. T. Hemann, E. HernandoMonge, D. Mu, S. Goodson,S. Powers, C. Cordon-Cardo, S. W. Lowe, G. J. Hannon and S. M. Hammond. 2005. A microRNA polycistron as a potential human oncogene. Nature 9, 828-833
  15. Hutvagner, G., J. Mcl.achlan, A. E. Pasquinelli, E. Balint, T. Tuschl and P. D. Zamore. 2001. A cellular function for the RNAinterference enzyme Dicer in the maturation of the let-7 small temporal RNA. Science 293, 834-838 https://doi.org/10.1126/science.1062961
  16. Ketting, R. F., S. E. Fischer, E. Bernstein, T. Sijen, G. J. Hannon and R. H. Plasterk. 2001. Dicer functions in RNA interference and in synthesis of small RNA involved in developmental timing in C. elegans. Genes Dev. 15, 2654-2659 https://doi.org/10.1101/gad.927801
  17. Knight, S. W. and B. L. Bass. 2001. A role for the RNase III enzyme DCR-1 in RNA interference and germ line development in Caenorhabditis elegans. Science 293, 2269-2271 https://doi.org/10.1126/science.1062039
  18. Landthaler, M, A. Yalcin and T. Tuschl. 2004. The human DiGeorge syndrome critical region gene 8 and Its D. melanogaster homolog are required for miRNA biogenesis. Curro BioI. 14, 2162-2167 https://doi.org/10.1016/j.cub.2004.11.001
  19. Lee, Y., C. Ahn, J. Han, H. Choi, J. Kim, J. Vim, J. Lee, P. Provost, O. Radmark, S. Kim and V. N. Kim. 2003. The nuclear RNase III Drosha initiates micro RNA processing. Nature 425, 415-419 https://doi.org/10.1038/nature01957
  20. Lund, E., S. Guttinger, A. Calado, J. E. Dahlberg and U. Kutay. 2004. Nuclear export of micro RNA precursors. Science 303, 95-98 https://doi.org/10.1126/science.1090599
  21. Miller, G., J. F. Enders, H. Lisco and H. I. Kohn. 1969. Establishment of lines from normal human blood leukocytes by co-cultivation with a leukocyte line derived from a leukemic child. Proc. Soc. Exp. Bioi. Med. 132, 247-252
  22. Miller, G. and M. Lipman. 1973. Release of infectious Epstein-Barr virus by transformed marmoset leukocytes. Proc. Natl. Acad. Sci. USA 70, 190-194
  23. Oudejans, J. J., A. J. van den Brule, N. M. Jiwa, P. C. de Bruin, G. J. Ossenkoppele, P. van der Valk, J. M Walboomers and C. J. Meijer. 1995. BHRF1, the Epstein-Barr virus (EBV) homologue of the BCL-2 protooncogene, is transcribed in EBV-associated B-celilymphomas and in reactive lymphocytes. Blood 86, 1893-1902
  24. Pfeffer,S., A. Sewer, M. Aagos-Quintana, R. Sheridan, C. Sander, F. A. Grasser, L. F. van Dyk, C. K. Ho, S. Shuman, M. Chien, J. J. Russo, J. Ju, G. Randall, B. D. Lindenbach, C. M Rice, V. Simon, D. D. Ho, M Zavolan and T. Tuschl. 2005. Identification of microRNAs of the herpesvirus family. Nat. Methods 2, 269-276 https://doi.org/10.1038/nmeth746
  25. Pfeffer, S., M. Zavolan, F. A Grasser, M. Chien, J. J. Russo, J. Ju, B. John, A J. Enright, D. Marks, C. Sander and T. Tuschl. 2004. Identification of virus-encoded microRNAs. Science 304, 734-736 https://doi.org/10.1126/science.1096781
  26. Provost, P., D. Dishart, J. Doucet, D. Frendewey, B. Samuelsson and O. Radmark. 2002. Ribonuclease activity and RNA binding of recombinant human Dicer. EMBO J. 21, 5864-5874 https://doi.org/10.1093/emboj/cdf578
  27. Rodriguez, A, S. Criffiths-Ioncs, J. L. Ashurst and A. Bradley. 2004. Identification of Mammalian microRNA Host Genes and Transcription Units. Genome Research 14, 1902-1910 https://doi.org/10.1101/gr.2722704
  28. Schmittgen, T. D., B. A Zakrajsek, A G. Mills, V. Gom, M. J. Singer and M. W. Reed. 2000. Quantitative reverse transcription-polymerase chain reaction to study mRNA decay: comparison of endpoint and real-time method. Anal. Biochem. 285, 194-204 https://doi.org/10.1006/abio.2000.4753
  29. Schmittgen, T. D., J. Jiang, Q. Liu and L. Yang. 2004. A high-throughput method to monitor the expression of micro RNA precursors. Nucleic Acids Res. 32, e43
  30. Smith, P. R., O. de Jesus, D. Turner, M. Hollyoake, C. E. Karstegl, B. E. Griffin, L. Karran, Y. Wang, S. D. Hayward and P. J. Farrell. 2000. Structure and coding content of CST (BART) family RNAs of Epstein-Barr virus. J. Virol. 74, 3082-3092 https://doi.org/10.1128/JVI.74.7.3082-3092.2000
  31. van Beek, J., A. A. Brink, M. B. Vervoort, M. J. van Zijp, C. J. Meijer, A. J. van den Brule and J. M. Middeldorp. 2003. In vivo transcription of the Epstein-Barr virus (EBV) BamHI-A region without associated in vivo BARFO protein expression in multiple EBV -associated disorders. J. Gen. Virol. 84, 2647-2659 https://doi.org/10.1099/vir.0.19196-0
  32. Yan, K. S., S. Yan, A Farooq, A Han, L. Zeng and M. M. Zhou. 2003. Structure and conserved RNA binding of the PAZ domain. Nature 426, 468-474 https://doi.org/10.1038/nature02129
  33. Yi, R., Y. Qin, I. G. Macara and B. R. Cullen. 2003. Exportin-5 mediates the nuclear export of pre-microRNAs and short hairpin RNAs. Genes Oev. 17, 3011-3016 https://doi.org/10.1101/gad.1158803