Abstract
Since the identification of the first virus-encoded microRNA (miRNA) in Epstein–Barr virus (EBV)-infected B cells in 2004, viral miRNAs have been found in different groups of herpesviruses. Viral miRNAs play an important role in regulating both viral and cellular gene expression. Identification and characterization of the cellular targets of viral miRNAs will not only advance our understanding of virus–cell interaction but might also reveal new strategies for the developments of antivirals. Our demonstration of the targeting of p53 upregulated modulator of apoptosis (PUMA) by an EBV-encoded viral miRNA provides one mechanism by which a viral miRNA facilitates viral replication by promoting cell survival. Using EBV miRNAs as an example, in this chapter, we detail the experimental procedures that can be used to identify and validate cellular targets of viral miRNAs.
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Abbreviations
- BART:
-
BamA rightward transcript
- CMV:
-
Cytomegalovirus
- EBV:
-
Epstein–Barr virus
- HSV-1:
-
Herpes simplex virus 1
- HSV-2:
-
Herpes simplex virus 2
- KSHV:
-
Kaposi’s sarcoma-associated herpesvirus
- NPC:
-
Nasopharyngeal carcinoma
- PUMA:
-
p53-upregulated modulator of apoptosis
- SV40:
-
Simian virus 40
- UTR:
-
Untranslated region
References
Ghosh, Z., Mallick, B., and Chakrabarti, J. (2009) Cellular versus viral microRNAs in host-virus interaction. Nucleic Acids Res 37, 1035–48.
Pfeffer, S., Zavolan, M., Grässer, F.A., Chien, M., Russo, J.J., Ju, J., John, B., Enright, A.J., Marks, D., Sander, C., and Tuschl, T. (2004) Identification of virus-encoded microRNAs. Science 304, 734–6.
Cullen, B.R. (2009) Viral and cellular messenger RNA targets of viral microRNAs. Nature 457, 421–5.
Choy, E.Y., Siu, K.L., Kok, K.H., Lung, R.W., Tsang, C.M., To, K.F., Kwong, D.L., Tsao, S.W., and Jin, D.Y. (2008) An Epstein-Barr virus-encoded microRNA targets PUMA to promote host cell survival. J Exp Med 27, 2551–60.
Cai, X., Schäfer, A., Lu, S., Bilello, J.P., Desrosiers, R.C., Edwards, R., Raab-Traub, N., and Cullen, B.R. (2006) Epstein-Barr virus microRNAs are evolutionarily conserved and differentially expressed. PLoS Pathog 2, e23.
Xia, T., O’Hara, A., Araujo, I., Barreto, J., Carvalho, E., Sapucaia, J.B., Ramos, J.C., Luz, E., Pedroso, C., Manrique, M., Toomey, N.L., Brites, C., Dittmer D.P., and Harrington, W.J. Jr. (2008) EBV microRNAs in primary lymphomas and targeting of CXCL-11 by ebv-mir-BHRF1-3. Cancer Res 68, 1436–42.
Cheung, S.T., Huang, D.P., Hui, A.B.Y., Lo, K.W., Ko, C.W., Tsang, Y.S., Wong, N., Whitney, B.M., and Lee, J.C.K. (1999) Nasopharyngeal carcinoma cell line (C666-1) consistently harbouring Epstein-Barr virus. Int J Cancer 83, 121–6.
Griffiths-Jones, S., Grocock, R.J., van Dongen, S., Bateman, A., and Enright, A.J. (2006) miRBase: microRNA sequences, targets and gene nomenclature. Nucleic Acids Res 34, D140–4.
Betel, D., Wilson, M., Gabow, A., Marks, D.S., and Sander, C. (2008) The microRNA.org resource: targets and expression. Nucleic Acids Res 36, D149–53.
Grimson, A., Farh, K.K., Johnston, W.K., Garrett-Engele, P., Lim, L.P., and Bartel D.P. (2005) MicroRNA targeting specificity in mammals: determinants beyond seed pairing. Mol Cell 27, 91–105.
Friedman, R.C., Farh, K.K., Burge, C.B., and Bartel, D.P. (2009) Most mammalian mRNAs are conserved targets of microRNAs. Genome Res 19, 92–105.
Bartel, D.P. (2009) MicroRNAs: target recognition and regulatory functions. Cell 136, 215–33.
Doench, J.G., Petersen, C.P., and Sharp, P.A. (2003) siRNAs can function as miRNAs. Genes Dev 17, 438–42.
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Kok, KH., Lei, T., Jin, DY. (2010). Identification and Validation of the Cellular Targets of Virus-Encoded MicroRNAs. In: Monticelli, S. (eds) MicroRNAs and the Immune System. Methods in Molecular Biology, vol 667. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-60761-811-9_21
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DOI: https://doi.org/10.1007/978-1-60761-811-9_21
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