DNA Viruses pp 231-244 | Cite as

Identification and Characterization of Herpesviral Immediate-Early Genes

  • Yan Yuan
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 292)

Abstract

Immediate-early (IE) genes are the first class of viral genes expressed after primary infection or reactivation. As transcription of IE genes does not require prior viral protein synthesis, this class of genes is experimentally defined by their transcription following primary infection or reactivation in the presence of inhibitors of protein synthesis. This chapter describes an approach to identify IE genes in a novel herpesvirus genome. Transcription of IE genes is selectively induced with sodium butyrate in the presence of the protein synthesis inhibitor cycloheximide. The transcripts of the induced genes are identified by using a cDNA subtraction-based method of gene expression screening.

Key Words

Immediate-early transcripts cDNA subtraction KSHV δ-herpesvirus reactivation 

References

  1. 1.
    Roizman, B. and Pellett, P. (2002) The family Herpesviridae: a brief introduction, in Fields Virology, 4th ed. (Knipe, D. M., Howley, P. M., Griffin, D. E., et al., eds.), Lippincott Williams & Wilkins, Philadelphia, pp. 2381–2397.Google Scholar
  2. 2.
    Miller, G. (1990) The switch between latency and replication of Epstein-Barr virus. J. Infect. Dis. 161, 833–844.PubMedGoogle Scholar
  3. 3.
    Wu, F. Y., Chen, H., Wang, S. E., et al. (2003) CCAAT/enhancer binding protein alpha interacts with ZTA and mediates ZTA-induced p21CIP-1 accumulation and G1 cell cycle arrest during the Epstein-Barr virus lytic cycle. J. Virol. 77, 1481–1500.PubMedCrossRefGoogle Scholar
  4. 4.
    Morrison, T. E., Mauser, A., Wong, A., Ting, J. P., and Kenney, S. C. (2001) Inhibition of IFN-gamma signaling by an Epstein-Barr virus immediate-early protein. Immunity 15, 787–799.PubMedCrossRefGoogle Scholar
  5. 5.
    Honess, R. W. and Roizman, B. (1974) Regulation of herpesvirus macromolecular synthesis. I. Cascade regulation of the synthesis of three groups of viral proteins. J. Virol. 14, 8–19.PubMedGoogle Scholar
  6. 6.
    Wathen, M. W., Thomsen, D. R., and Stinski, M. F. (1981) Temporal regulation of human cytomegalovirus transcription at immediate early and early times after infection. J. Virol. 38, 446–459.PubMedGoogle Scholar
  7. 7.
    Zhu, F. X., Cusano, T., and Yuan, Y. (1999) Identification of the immediate-early transcripts of Kaposi’s sarcoma-associated herpesvirus. J. Virol. 73, 5556–5567.PubMedGoogle Scholar
  8. 8.
    Ebrahimi, B., Dutia, B. M., Roberts, K. L., et al. (2003) Transcriptome profile of murine gammaherpesvirus-68 lytic infection. J. Gen. Virol. 84, 99–109.PubMedCrossRefGoogle Scholar
  9. 9.
    Cesarman, E., Moore, P. S., Rao, P. H., Inghirami, G., Knowles, D. M., and Chang, Y. (1995) In vitro establishment and characterization of two acquired immunodeficiency syndrome-related lymphoma cell lines (BC-1 and BC-2) containing Kaposi’s sarcoma-associated herpesvirus-like (KSHV) DNA sequences. Blood 86, 2708–2714.PubMedGoogle Scholar
  10. 10.
    Renne, R., Zhong, W., Herndier, B., et al. (1996) Lytic growth of Kaposi’s sarcoma-associated herpesvirus (human herpesvirus 8) in culture. Nat. Med. 2,342–346.CrossRefGoogle Scholar
  11. 11.
    Wang, Z. and Brown, D. D. (1991) A gene expression screen. Proc. Natl.Acad. Sci. USA 88, 11505–11509.PubMedCrossRefGoogle Scholar

Copyright information

© Humana Press Inc. 2005

Authors and Affiliations

  • Yan Yuan
    • 1
  1. 1.Department of MicrobiologyUniversity of Pennsylvania School of Dental MedicinePhiladelphia

Personalised recommendations