Skip to main content
NCBI home page
Molecular and Cellular Biology logoLink to Molecular and Cellular Biology
. 1997 Apr;17(4):2207–2216. doi: 10.1128/mcb.17.4.2207

Sequence-mediated regulation of adenovirus gene expression by repression of mRNA accumulation.

J C Prescott 1, L Liu 1, E Falck-Pedersen 1
PMCID: PMC232070  PMID: 9121471

Abstract

Gene expression in complex transcription units can be regulated at virtually every step in the production of mature cytoplasmic mRNA, including transcription initiation, elongation, termination, pre-mRNA processing, nucleus-to-cytoplasm mRNA transport, and alterations in mRNA stability. We have been characterizing alternative poly(A) site usage in the adenovirus major late transcription unit (MLTU) as a model for regulation at the level of pre-mRNA 3'-end processing. The MLTU contains five polyadenylation sites (L1 through L5). The promoter proximal site (L1) functions as the dominant poly(A) site during the early stage of adenovirus infection and in plasmid transfections when multiple poly(A) sites are present at the 3' end of a reporter plasmid. In contrast, stable mRNA processed at all five poly(A) sites is found during the late stage of adenovirus infection, after viral DNA replication has begun. Despite its dominance during early infection, L1 is a comparatively poor substrate for 3'-end RNA processing both in vivo and in vitro. In this study we have investigated the basis for the early L1 dominance. We have found that mRNA containing an unprocessed L1 poly(A) site is compromised in its ability to enter the steady-state pool of stable mRNA. This inhibition, which affects either the nuclear stability or nucleus-to-cytoplasm transport of the pre-mRNA, requires a cis-acting sequence located upstream of the L1 poly(A) site.

Full Text

The Full Text of this article is available as a PDF (808.2 KB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Batt D. B., Luo Y., Carmichael G. G. Polyadenylation and transcription termination in gene constructs containing multiple tandem polyadenylation signals. Nucleic Acids Res. 1994 Jul 25;22(14):2811–2816. doi: 10.1093/nar/22.14.2811. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bhat B. M., Wold W. S. ATTAAA as well as downstream sequences are required for RNA 3'-end formation in the E3 complex transcription unit of adenovirus. Mol Cell Biol. 1985 Nov;5(11):3183–3193. doi: 10.1128/mcb.5.11.3183. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Brasier A. R., Tate J. E., Habener J. F. Optimized use of the firefly luciferase assay as a reporter gene in mammalian cell lines. Biotechniques. 1989 Nov-Dec;7(10):1116–1122. [PubMed] [Google Scholar]
  4. Chomczynski P., Sacchi N. Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Anal Biochem. 1987 Apr;162(1):156–159. doi: 10.1006/abio.1987.9999. [DOI] [PubMed] [Google Scholar]
  5. Chow L. T., Gelinas R. E., Broker T. R., Roberts R. J. An amazing sequence arrangement at the 5' ends of adenovirus 2 messenger RNA. Cell. 1977 Sep;12(1):1–8. doi: 10.1016/0092-8674(77)90180-5. [DOI] [PubMed] [Google Scholar]
  6. Citron B., Falck-Pedersen E., Salditt-Georgieff M., Darnell J. E., Jr Transcription termination occurs within a 1000 base pair region downstream from the poly(A) site of the mouse beta-globin (major) gene. Nucleic Acids Res. 1984 Nov 26;12(22):8723–8731. doi: 10.1093/nar/12.22.8723. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. DeZazzo J. D., Falck-Pedersen E., Imperiale M. J. Sequences regulating temporal poly(A) site switching in the adenovirus major late transcription unit. Mol Cell Biol. 1991 Dec;11(12):5977–5984. doi: 10.1128/mcb.11.12.5977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. DeZazzo J. D., Imperiale M. J. Sequences upstream of AAUAAA influence poly(A) site selection in a complex transcription unit. Mol Cell Biol. 1989 Nov;9(11):4951–4961. doi: 10.1128/mcb.9.11.4951. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Denome R. M., Cole C. N. Patterns of polyadenylation site selection in gene constructs containing multiple polyadenylation signals. Mol Cell Biol. 1988 Nov;8(11):4829–4839. doi: 10.1128/mcb.8.11.4829. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Edwalds-Gilbert G., Prescott J., Falck-Pedersen E. 3' RNA processing efficiency plays a primary role in generating termination-competent RNA polymerase II elongation complexes. Mol Cell Biol. 1993 Jun;13(6):3472–3480. doi: 10.1128/mcb.13.6.3472. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Falck-Pedersen E., Heinflink M., Alvira M., Nussenzveig D. R., Gershengorn M. C. Expression of thyrotropin-releasing hormone receptors by adenovirus-mediated gene transfer reveals that thyrotropin-releasing hormone desensitization is cell specific. Mol Pharmacol. 1994 Apr;45(4):684–689. [PubMed] [Google Scholar]
  12. Falck-Pedersen E., Logan J. Regulation of poly(A) site selection in adenovirus. J Virol. 1989 Feb;63(2):532–541. doi: 10.1128/jvi.63.2.532-541.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Falck-Pedersen E., Logan J., Shenk T., Darnell J. E., Jr Transcription termination within the E1A gene of adenovirus induced by insertion of the mouse beta-major globin terminator element. Cell. 1985 Apr;40(4):897–905. doi: 10.1016/0092-8674(85)90349-6. [DOI] [PubMed] [Google Scholar]
  14. Galli G., Guise J. W., McDevitt M. A., Tucker P. W., Nevins J. R. Relative position and strengths of poly(A) sites as well as transcription termination are critical to membrane versus secreted mu-chain expression during B-cell development. Genes Dev. 1987 Jul;1(5):471–481. doi: 10.1101/gad.1.5.471. [DOI] [PubMed] [Google Scholar]
  15. Gilmartin G. M., Hung S. L., DeZazzo J. D., Fleming E. S., Imperiale M. J. Sequences regulating poly(A) site selection within the adenovirus major late transcription unit influence the interaction of constitutive processing factors with the pre-mRNA. J Virol. 1996 Mar;70(3):1775–1783. doi: 10.1128/jvi.70.3.1775-1783.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Gilmartin G. M., McDevitt M. A., Nevins J. R. Multiple factors are required for specific RNA cleavage at a poly(A) addition site. Genes Dev. 1988 May;2(5):578–587. doi: 10.1101/gad.2.5.578. [DOI] [PubMed] [Google Scholar]
  17. Graham F. L., Smiley J., Russell W. C., Nairn R. Characteristics of a human cell line transformed by DNA from human adenovirus type 5. J Gen Virol. 1977 Jul;36(1):59–74. doi: 10.1099/0022-1317-36-1-59. [DOI] [PubMed] [Google Scholar]
  18. Hales K. H., Birk J. M., Imperiale M. J. Analysis of adenovirus type 2 L1 RNA 3'-end formation in vivo and in vitro. J Virol. 1988 Apr;62(4):1464–1468. doi: 10.1128/jvi.62.4.1464-1468.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Iwamoto S., Eggerding F., Falck-Pederson E., Darnell J. E., Jr Transcription unit mapping in adenovirus: regions of termination. J Virol. 1986 Jul;59(1):112–119. doi: 10.1128/jvi.59.1.112-119.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Jiménez-García L. F., Spector D. L. In vivo evidence that transcription and splicing are coordinated by a recruiting mechanism. Cell. 1993 Apr 9;73(1):47–59. doi: 10.1016/0092-8674(93)90159-n. [DOI] [PubMed] [Google Scholar]
  21. Jones N., Shenk T. Isolation of adenovirus type 5 host range deletion mutants defective for transformation of rat embryo cells. Cell. 1979 Jul;17(3):683–689. doi: 10.1016/0092-8674(79)90275-7. [DOI] [PubMed] [Google Scholar]
  22. Le Moullec J. M., Akusjärvi G., Stålhandske P., Pettersson U., Chambraud B., Gilardi P., Nasri M., Perricaudet M. Polyadenylic acid addition sites in the adenovirus type 2 major late transcription unit. J Virol. 1983 Oct;48(1):127–134. doi: 10.1128/jvi.48.1.127-134.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Levitt N., Briggs D., Gil A., Proudfoot N. J. Definition of an efficient synthetic poly(A) site. Genes Dev. 1989 Jul;3(7):1019–1025. doi: 10.1101/gad.3.7.1019. [DOI] [PubMed] [Google Scholar]
  24. Nevins J. R., Chen-Kiang S. Processing of adenovirus nuclear RNA to mRNA. Adv Virus Res. 1981;26:1–35. doi: 10.1016/s0065-3527(08)60419-4. [DOI] [PubMed] [Google Scholar]
  25. Nevins J. R., Darnell J. E. Groups of adenovirus type 2 mRNA's derived from a large primary transcript: probable nuclear origin and possible common 3' ends. J Virol. 1978 Mar;25(3):811–823. doi: 10.1128/jvi.25.3.811-823.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Nevins J. R., Wilson M. C. Regulation of adenovirus-2 gene expression at the level of transcriptional termination and RNA processing. Nature. 1981 Mar 12;290(5802):113–118. doi: 10.1038/290113a0. [DOI] [PubMed] [Google Scholar]
  27. Prescott J. C., Falck-Pedersen E. Varied poly(A) site efficiency in the adenovirus major late transcription unit. J Biol Chem. 1992 Apr 25;267(12):8175–8181. [PubMed] [Google Scholar]
  28. Prescott J., Falck-Pedersen E. Sequence elements upstream of the 3' cleavage site confer substrate strength to the adenovirus L1 and L3 polyadenylation sites. Mol Cell Biol. 1994 Jul;14(7):4682–4693. doi: 10.1128/mcb.14.7.4682. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Puvion-Dutilleul F., Puvion E. Replicating single-stranded adenovirus type 5 DNA molecules accumulate within well-delimited intranuclear areas of lytically infected HeLa cells. Eur J Cell Biol. 1990 Aug;52(2):379–388. [PubMed] [Google Scholar]
  30. Shaw A. R., Ziff E. B. Transcripts from the adenovirus-2 major late promoter yield a single early family of 3' coterminal mRNAs and five late families. Cell. 1980 Dec;22(3):905–916. doi: 10.1016/0092-8674(80)90568-1. [DOI] [PubMed] [Google Scholar]
  31. Stow N. D. Cloning of a DNA fragment from the left-hand terminus of the adenovirus type 2 genome and its use in site-directed mutagenesis. J Virol. 1981 Jan;37(1):171–180. doi: 10.1128/jvi.37.1.171-180.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Tantravahi J., Alvira M., Falck-Pedersen E. Characterization of the mouse beta maj globin transcription termination region: a spacing sequence is required between the poly(A) signal sequence and multiple downstream termination elements. Mol Cell Biol. 1993 Jan;13(1):578–587. doi: 10.1128/mcb.13.1.578. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Weiss E. A., Gilmartin G. M., Nevins J. R. Poly(A) site efficiency reflects the stability of complex formation involving the downstream element. EMBO J. 1991 Jan;10(1):215–219. doi: 10.1002/j.1460-2075.1991.tb07938.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Wilson-Gunn S. I., Kilpatrick J. E., Imperiale M. J. Regulated adenovirus mRNA 3'-end formation in a coupled in vitro transcription-processing system. J Virol. 1992 Sep;66(9):5418–5424. doi: 10.1128/jvi.66.9.5418-5424.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Molecular and Cellular Biology are provided here courtesy of Taylor & Francis

RESOURCES