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. 1982 Oct 11;10(19):6177–6190. doi: 10.1093/nar/10.19.6177

Effect of intron size on splicing efficiency in retroviral transcripts.

B R Cullen, J J Kopchick, D W Stacey
PMCID: PMC320959  PMID: 6292870

Abstract

To study the effect of intron size on splicing efficiency we have varied the size of the avian leukosis virus (ALV) env mRNA intron in a cloned ALV genome. This was accomplished by deletion of ALV sequences or insertion of phage lambda DNA. The effect of these modifications on splicing was analyzed by microinjection of the modified clones into RSV(-) chicken cells. Viral env mRNA when transcribed and properly spliced within these cells complemented the RSV(-) env deficiency leading to the production of focus forming units. Using this assay it was shown that deletion of up to 3.7 kb of the 4.68 kb env intron did not inhibit correct splicing nor did insertion of up to 8 kb of phage lambda DNA prevent splicing. Our results indicate that intron size can be varied over a wide range without preventing splicing.

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Selected References

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

  1. Coffin J. M. Structure, replication, and recombination of retrovirus genomes: some unifying hypotheses. J Gen Virol. 1979 Jan;42(1):1–26. doi: 10.1099/0022-1317-42-1-1. [DOI] [PubMed] [Google Scholar]
  2. Gilbert W. Why genes in pieces? Nature. 1978 Feb 9;271(5645):501–501. doi: 10.1038/271501a0. [DOI] [PubMed] [Google Scholar]
  3. Gruss P., Efstratiadis A., Karathanasis S., König M., Khoury G. Synthesis of stable unspliced mRNA from an intronless simian virus 40--rat preproinsulin gene recombinant. Proc Natl Acad Sci U S A. 1981 Oct;78(10):6091–6095. doi: 10.1073/pnas.78.10.6091. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Hackett P. B., Swanstrom R., Varmus H. E., Bishop J. M. The leader sequence of the subgenomic mRNA's of Rous sarcoma virus is approximately 390 nucleotides. J Virol. 1982 Feb;41(2):527–534. doi: 10.1128/jvi.41.2.527-534.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Hamer D. H., Leder P. Splicing and the formation of stable RNA. Cell. 1979 Dec;18(4):1299–1302. doi: 10.1016/0092-8674(79)90240-x. [DOI] [PubMed] [Google Scholar]
  6. Ju G., Boone L., Skalka A. M. Isolation and characterization of recombinant DNA clones of avian retroviruses: size heterogeneity and instability of the direct repeat. J Virol. 1980 Mar;33(3):1026–1033. doi: 10.1128/jvi.33.3.1026-1033.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Khoury G., Gruss P., Dhar R., Lai C. J. Processing and expression of early SV40 mRNA: a role for RNA conformation in splicing. Cell. 1979 Sep;18(1):85–92. doi: 10.1016/0092-8674(79)90356-8. [DOI] [PubMed] [Google Scholar]
  8. Kopchick J. J., Ju G., Skalka A. M., Stacey D. W. Biological activity of cloned retroviral DNA in microinjected cells. Proc Natl Acad Sci U S A. 1981 Jul;78(7):4383–4387. doi: 10.1073/pnas.78.7.4383. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Lomedico P., Rosenthal N., Efstratidadis A., Gilbert W., Kolodner R., Tizard R. The structure and evolution of the two nonallelic rat preproinsulin genes. Cell. 1979 Oct;18(2):545–558. doi: 10.1016/0092-8674(79)90071-0. [DOI] [PubMed] [Google Scholar]
  10. Mulligan R. C., Berg P. Factors governing the expression of a bacterial gene in mammalian cells. Mol Cell Biol. 1981 May;1(5):449–459. doi: 10.1128/mcb.1.5.449. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Nevins J. R. Adenovirus gene expression: control at multiple steps of mRNA biogenesis. Cell. 1982 Jan;28(1):1–2. doi: 10.1016/0092-8674(82)90366-x. [DOI] [PubMed] [Google Scholar]
  12. Ohkubo H., Vogeli G., Mudryj M., Avvedimento V. E., Sullivan M., Pastan I., de Crombrugghe B. Isolation and characterization of overlapping genomic clones covering the chicken alpha 2 (type I) collagen gene. Proc Natl Acad Sci U S A. 1980 Dec;77(12):7059–7063. doi: 10.1073/pnas.77.12.7059. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Perry R. P., Kelley D. E. Immunoglobulin messenger RNAs in murine cell lines that have characteristics of immature B lymphocytes. Cell. 1979 Dec;18(4):1333–1339. doi: 10.1016/0092-8674(79)90243-5. [DOI] [PubMed] [Google Scholar]
  14. Rosenberg M., Court D. Regulatory sequences involved in the promotion and termination of RNA transcription. Annu Rev Genet. 1979;13:319–353. doi: 10.1146/annurev.ge.13.120179.001535. [DOI] [PubMed] [Google Scholar]
  15. Shimotohno K., Temin H. M. Formation of infectious progeny virus after insertion of herpes simplex thymidine kinase gene into DNA of an avian retrovirus. Cell. 1981 Oct;26(1 Pt 1):67–77. doi: 10.1016/0092-8674(81)90034-9. [DOI] [PubMed] [Google Scholar]
  16. Stacey D. W., Allfrey V. G., Hanafusa H. Microinjection analysis of envelope-glycoprotein messenger activities of avian leukosis viral RNAs. Proc Natl Acad Sci U S A. 1977 Apr;74(4):1614–1618. doi: 10.1073/pnas.74.4.1614. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Stacey D. W. Expression of a subgenomic retroviral messenger RNA. Cell. 1980 Oct;21(3):811–820. doi: 10.1016/0092-8674(80)90444-4. [DOI] [PubMed] [Google Scholar]
  18. Stacey D. W., Hanafusa H. Nuclear conversion of microinjected avian leukosis virion RNA into an envelope-glycoprotein messenger. Nature. 1978 Jun 29;273(5665):779–782. doi: 10.1038/273779a0. [DOI] [PubMed] [Google Scholar]
  19. Swanstrom R., Varmus H. E., Bishop J. M. Nucleotide sequence of the 5' noncoding region and part of the gag gene of Rous sarcoma virus. J Virol. 1982 Feb;41(2):535–541. doi: 10.1128/jvi.41.2.535-541.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Tiemeier D. C., Tilghman S. M., Polsky F. I., Seidman J. G., Leder A., Edgell M. H., Leder P. A comparison of two cloned mouse beta-globin genes and their surrounding and intervening sequences. Cell. 1978 Jun;14(2):237–245. doi: 10.1016/0092-8674(78)90110-1. [DOI] [PubMed] [Google Scholar]
  21. Wang L. H. The gene order of avian RNA tumor viruses derived from biochemical analyses of deletion mutants and viral recombinants. Annu Rev Microbiol. 1978;32:561–592. doi: 10.1146/annurev.mi.32.100178.003021. [DOI] [PubMed] [Google Scholar]
  22. Wei C. M., Gibson M., Spear P. G., Scolnick E. M. Construction and isolation of a transmissible retrovirus containing the src gene of Harvey murine sarcoma virus and the thymidine kinase gene of herpes simplex virus type 1. J Virol. 1981 Sep;39(3):935–944. doi: 10.1128/jvi.39.3.935-944.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Young R. A., Hagenbüchle O., Schibler U. A single mouse alpha-amylase gene specifies two different tissue-specific mRNAs. Cell. 1981 Feb;23(2):451–458. doi: 10.1016/0092-8674(81)90140-9. [DOI] [PubMed] [Google Scholar]
  24. van den Berg J., van Ooyen A., Mantei N., Schamböck A., Grosveld G., Flavell R. A., Weissmann C. Comparison of cloned rabbit and mouse beta-globin genes showing strong evolutionary divergence of two homologous pairs of introns. Nature. 1978 Nov 2;276(5683):37–44. doi: 10.1038/276037a0. [DOI] [PubMed] [Google Scholar]

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