Skip to main content
SpringerLink
Log in

A Role for RNA Processing in Regulating Expression from Transfected Genes

  • Published:
Somatic Cell and Molecular Genetics

Abstract

We have examined the expression of cloned genes following their stable integration into the genome of pluripotent embryonal carcinoma stem cells. Transfected genes integrate into the genome as tandem arrays. Expression of reporter genes from these tandem arrays in embryonal carcinoma cells is inefficient probably because genes are subject to repeat-induced gene silencing. We found that expression of reporter genes was significantly enhanced if co-transfected with cloned fragments derived from the murine Pgk-1 gene. The enhanced expression required (a) that the Pgk-1 fragment carries an active promoter, (b) that the promoter drives transcription through a region of more than 12 kbp, and (c) that this transcribed region contains both introns and exons. Reporter gene activity did not require specific Pgk-1 DNA sequences suggesting that the coupled processes of transcription and RNA processing conferred activity on neighboring genes probably by influencing local chromatin structure. Consistent with this idea, the effect of the Pgk-1 gene could be mimicked by exposing cells to butyrate or trichostatin A, inhibitors of histone deacetylase. Thus, the effect of the co-transfected Pgk-1 gene is to inhibit the process of gene inactivation possibly by functioning like an insulator or boundary element in the chromatin.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

LITERATURE CITED

  1. Wigler, M., Sweet, R., Sim, G.K., Wold, B., Pellicer, A., Lacy, E., Maniatis, T., Silverstein, S., and Axel, R. (1977). Cell 16:777–785.

    Google Scholar 

  2. Meyer, P. (1996). Biol. Chem. Hoppe Seyler 377:87–95.

    PubMed  Google Scholar 

  3. Dorer, D.R., and Henikoff, S. (1994). Cell 77:993–1002.

    Article  PubMed  Google Scholar 

  4. Pal-Bhadra, M., Bhadra, U., and Birchler, J.A. (1997). Cell 90:479–490.

    PubMed  Google Scholar 

  5. Sabl, J.F., and Henikoff, S. (1996). Genetics 142:447–458.

    PubMed  Google Scholar 

  6. Singer, M.J., and Selker, E.U. (1995). Curr. Top. Microbiol. Immunol. 197:165–178.

    PubMed  Google Scholar 

  7. Dougherty, W.G., and Parks, T.D. (1995). Curr. Opin. Cell Biol. 7:399–405.

    PubMed  Google Scholar 

  8. Meyer, P., and Saedler, H. (1996). Annu. Rev. Plant Physiol. Plant Mol. Biol. 47:23–48.

    PubMed  Google Scholar 

  9. Irelan, J.T., and Selker, E.U. (1997). Genetics 146:509–523.

    PubMed  Google Scholar 

  10. Garrick, D., Fiering, S., Martin, D.I.K., and Whitelaw, E. (1998). Nature Genet. 18:56–59.

    PubMed  Google Scholar 

  11. Gerasimova, T.I., and Corces, V.G. (1996). Curr. Opin. Genet. Dev. 6:185–192.

    PubMed  Google Scholar 

  12. McBurney, M.W., Fournier, S., Schmidt-Kastner, P.K., Jardine, K., and Craig, J. (1995). Somatic Cell Mol. Genet. 20:529–540.

    Google Scholar 

  13. Schmidt-Kastner, P.K., Jardine, K., Cormier, M., and McBurney, M.W. (1996). Somat. Cell Mol. Genet. 22:383–392.

    PubMed  Google Scholar 

  14. McBurney, M.W., Fournier, S., Jardine, K., and Sutherland, L. (1995). Somatic Cell Mol. Genet. 20:515–528.

    Google Scholar 

  15. McBurney, M.W., Sutherland, L.C., Adra, C.N., Leclair, B., Rudnicki, M.A., and Jardine, K. (1991). Nucleic Acids Res. 19:5755–5761.

    PubMed  Google Scholar 

  16. Sutherland, L.C., St-Arnaud, R., and McBurney, M.W. (1995). Gene Expr. 4:265–279.

    PubMed  Google Scholar 

  17. McBurney, M.W., and Rogers, B.J. (1982). Dev. Biol. 89:503–508.

    PubMed  Google Scholar 

  18. Rudnicki, M.A., and McBurney, M.W. (1987). In: Teratocarcinomas and embryonic stem cells, a practical approach. E.J. Robertson, editor. IRL Press, Oxford. 19–49.

    Google Scholar 

  19. Chen, C., and Okayama, H. (1987). Mol. Cell. Biol. 7:2745–2752.

    PubMed  Google Scholar 

  20. Birnboim, H.C., and Doly, J. (1979). Nucleic Acids Res. 7:1513–1520.

    PubMed  Google Scholar 

  21. Adra, C.N., Boer, P.H., and McBurney, M.W. (1987). Gene 60:65–74.

    PubMed  Google Scholar 

  22. Boer, P.H., Potten, H., Adra, C.N., Jardine, K., Mullhofer, G., and McBurney, M.W. (1990). Biochem. Genet. 28:299–308.

    PubMed  Google Scholar 

  23. Hannan, G.N., Lehnert, S.A., MacAvoy, E.S., Jennings, P.A., and Molloy, P.L. (1993). Gene 130:233–239.

    PubMed  Google Scholar 

  24. Adra, C.N., Ellis, N.A., and McBurney, M.W. (1988). Somatic Cell Mol. Genet. 14:69–81.

    PubMed  Google Scholar 

  25. Sambrook, J., Fritsch, E.F., and Maniatis, T. (1989). Molecular Cloning, A Laboratory Manual. Harbor Laboratory Press, Cold Spring Harbor, New York.

    Google Scholar 

  26. McBurney, M.W., and Strutt, B.J. (1980). Cell 21:357–364.

    PubMed  Google Scholar 

  27. Di Fruscio, M., Gilchrist, C.A., Baker, R.T., and Gray, D.A. (1998). Biochim. Biophys. Acta 1398:9–17.

    PubMed  Google Scholar 

  28. Coffey, A.J., Roberts, R.G., Green, E.D., Cole, C.G., Butler, R., Anand, R., Giannelli, F., and Bentley, D.R. (1992). Genomics 12:474–484.

    PubMed  Google Scholar 

  29. Michaelson, A.M., Blake, C.C.F., Evans, S.T., and Orkin, S.H. (1985). Proc. Natl. Acad. Sci. U.S.A. 82:6965–6969.

    PubMed  Google Scholar 

  30. Norton, P.A., and Coffin, J.M. (1985). Mol. Cell. Biol. 5:281–290.

    PubMed  Google Scholar 

  31. Laird, P.W., Zidjerveld, A., Linders, K., Rudnicki, M.A., Jaenisch, R., and Berns, A. (1991). Nucleic Acids Res. 19:4293.

    PubMed  Google Scholar 

  32. Johnson, P., and Friedmann, T. (1990). Gene 88:207–213.

    PubMed  Google Scholar 

  33. Prado, F., Piruat, J.I., and Aguilera, A. (1997). EMBO J. 16:2826–2835.

    PubMed  Google Scholar 

  34. Grey, M., Düsterhöft, A., Henriques, J.A.P., and Brendel, M. (1996). Nucleic Acids Res. 24:4009–4014.

    PubMed  Google Scholar 

  35. Dorer, D.R., and Henikoff, S. (1997). Genetics 147:1181–1190.

    PubMed  Google Scholar 

  36. Grunstein, M. (1997). Nature 389:349–352.

    PubMed  Google Scholar 

  37. Wade, P.A., Pruss, D., and Wolffe, A.P. (1997). Trends. Biochem. Sci. 22:1–132.

    PubMed  Google Scholar 

  38. Pazin, M.J., and Kadonaga, J.T. (1997). Cell 89:325–328.

    PubMed  Google Scholar 

  39. Candido, E.P., Reeves, R., and Davie, J.R. (1978). Cell 14:105–113.

    PubMed  Google Scholar 

  40. Yoshida, M., Kijima, M., Akita, M., and Beppu, T. (1990). J. Biol. Chem. 265:17174–17179.

    PubMed  Google Scholar 

  41. Chen, W.Y., Bailey, E.C., McCune, S.L., Dong, J.Y., and Townes, T.M. (1997). Proc. Natl. Acad. Sci. U.S.A. 94:5798–5803.

    PubMed  Google Scholar 

  42. Schmidt-Kastner, P.K., Jardine, K., Cormier, M., and McBurney, M.W. (1998). Oncogene 16:3003–3011.

    PubMed  Google Scholar 

  43. Mummenbrauer, T., Janus, F., Müller, B., Wiesmüller, L., Deppert, W., and Grosse, F. (1996). Cell 85:1089–1099.

    PubMed  Google Scholar 

  44. Bakalkin, G., Yakovleva, T., Selivanova, G., Magnusson, K.P., Szekely, L., Kiseleva, E., Klein, G., Terenius, L., and Wiman, K.G. (1994). Proc. Natl. Acad. Sci. U.S.A. 91:413–417.

    PubMed  Google Scholar 

  45. Wu, L., Bayle, J.H., Elenbaas, B., Pavletich, N.P., and Levine, A.J. (1995). Mol. Cell. Biol. 15:497–504.

    PubMed  Google Scholar 

  46. Stürzbecher, H.W., Donzelmann, B., Henning, W., Knippschild, U., and Buchhop, S. (1996). EMBO J. 15:1992–2002.

    PubMed  Google Scholar 

  47. Wiesmüller, L., Cammenga, J., and Deppert, W.W. (1996). J. Virol. 70:737–744.

    PubMed  Google Scholar 

  48. Mekeel, K.L., Tang, W., Kachnic, L.A., Luo, C.M., DeFrank, J.S., and Powell, S.N. (1997). Oncogene 14:1847–1857.

    PubMed  Google Scholar 

  49. Chuang, L.S.H., Ian, H.I., Koh, T.W., Ng, H.H., Xu, G.L., and Li, B.F.L. (1997). Science 277:1996–2000.

    PubMed  Google Scholar 

  50. Singer, R.H., and Green, M.R. (1997). Cell 91:291–294.

    PubMed  Google Scholar 

  51. Hertel, K.J., Lynch, K.W., and Maniatis, T. (1997). Curr. Opin. Cell Biol. 9:350–357.

    PubMed  Google Scholar 

  52. McCracken, S., Fong, N., Yankulov, K., Ballantyne, S., Pan, G.H., Greenblatt, J., Patterson, S.D., Wickens, M., and Bentley, D.L. (1997). Nature 385:357–361.

    PubMed  Google Scholar 

  53. Steinmetz, E.J. (1997). Cell 89:491–494.

    PubMed  Google Scholar 

  54. Greenblatt, J. (1997). Curr. Opin. Cell Biol. 9:310–319.

    PubMed  Google Scholar 

  55. Roth, S.Y., and Allis, C.D. (1996). Cell 87:5–8.

    PubMed  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Ottawa Regional Cancer Center and Department of Medicine, University of Ottawa, 501 Smyth Road, Ottawa, Canada, K1H 8L6

    Michael W. McBurney, Xiaofeng Yang, Karen Jardine & Michelle Cormier

Authors
  1. Michael W. McBurney
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Xiaofeng Yang
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Karen Jardine
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Michelle Cormier
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

McBurney, M.W., Yang, X., Jardine, K. et al. A Role for RNA Processing in Regulating Expression from Transfected Genes. Somat Cell Mol Genet 24, 203–215 (1998). https://doi.org/10.1023/B:SCAM.0000007123.70630.40

Download citation

  • Issue Date:

  • DOI: https://doi.org/10.1023/B:SCAM.0000007123.70630.40

Keywords

Search

Navigation