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. 1990 Nov;64(11):5260–5269. doi: 10.1128/jvi.64.11.5260-5269.1990

Activation of cryptic splice sites in murine sarcoma virus-124 mutants.

M de Mars 1, P E Cizdziel 1, E C Murphy Jr 1
PMCID: PMC248562  PMID: 2170671

Abstract

We have examined splice site activation in relation to intron structure in murine sarcoma virus (MuSV)-124 RNA. MuSV-124 contains inactive murine leukemia virus env gene splice sites (termed 5' env and 3' env) as well as cryptic sites in the gag and v-mos genes (termed 5' gag and 3' mos) which are activated for thermosensitive splicing by a 1,487-base intronic deletion in the MuSV-124 derived MuSVts110 retrovirus. To determine conditions permissive for splice site activation, we examined MuSV-124 mutants deleted in the 1,919-base intron bounded by the 5' gag and 3' mos sites. Several of these deletions activated thermosensitive splicing either at the same sites used in MuSVts110 or in a previously unreported temperature-sensitive splice event between the 5' gag and 3' env sites. These data suggested that the thermosensitive splicing phenotype characteristic of MuSVts110 required neither a specialized intron nor selection of a particular 3' splice site. The 3' env and 3' mos sites were found to compete for splicing to the 5' gag site; the more upstream 3' env site was exclusively used in MuSV-124 mutants containing both sites, whereas selection of the 3' mos site required removal of the 3' env site. Branchpoint sequences were found to have a potential regulatory role in thermosensitive splicing. Insertion of a beta-globin branchpoint sequence in a splicing-inactive MuSV-124 mutant activated efficient nonthermosensitive splicing at the 3' mos site, whereas a mutated branchpoint activated less efficient but thermosensitive 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. Arrigo S., Beemon K. Regulation of Rous sarcoma virus RNA splicing and stability. Mol Cell Biol. 1988 Nov;8(11):4858–4867. doi: 10.1128/mcb.8.11.4858. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Biggart N. W., Gallick G. E., Murphy E. C., Jr Nickel-induced heritable alterations in retroviral transforming gene expression. J Virol. 1987 Aug;61(8):2378–2388. doi: 10.1128/jvi.61.8.2378-2388.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Blair D. G., Hull M. A., Finch E. A. The isolation and preliminary characterization of temperature-sensitive transformation mutants of Moloney sarcoma virus. Virology. 1979 Jun;95(2):303–316. doi: 10.1016/0042-6822(79)90486-0. [DOI] [PubMed] [Google Scholar]
  4. Cellini A., Felder E., Rossi J. J. Yeast pre-messenger RNA splicing efficiency depends on critical spacing requirements between the branch point and 3' splice site. EMBO J. 1986 May;5(5):1023–1030. doi: 10.1002/j.1460-2075.1986.tb04317.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Chang D. D., Sharp P. A. Regulation by HIV Rev depends upon recognition of splice sites. Cell. 1989 Dec 1;59(5):789–795. doi: 10.1016/0092-8674(89)90602-8. [DOI] [PubMed] [Google Scholar]
  6. Chebli K., Gattoni R., Schmitt P., Hildwein G., Stevenin J. The 216-nucleotide intron of the E1A pre-mRNA contains a hairpin structure that permits utilization of unusually distant branch acceptors. Mol Cell Biol. 1989 Nov;9(11):4852–4861. doi: 10.1128/mcb.9.11.4852. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Cizdziel P. E., de Mars M., Murphy E. C., Jr Exploitation of a thermosensitive splicing event to study pre-mRNA splicing in vivo. Mol Cell Biol. 1988 Apr;8(4):1558–1569. doi: 10.1128/mcb.8.4.1558. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Cullen B. R., Kopchick J. J., Stacey D. W. Effect of intron size on splicing efficiency in retroviral transcripts. Nucleic Acids Res. 1982 Oct 11;10(19):6177–6190. doi: 10.1093/nar/10.19.6177. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. De Mars M., Sterner D. A., Chiocca S. M., Biggart N. W., Murphy E. C., Jr Regulation of RNA splicing in gag-deficient mutants of Moloney murine sarcoma virus MuSVts110. J Virol. 1990 Apr;64(4):1421–1428. doi: 10.1128/jvi.64.4.1421-1428.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Furdon P. J., Kole R. The length of the downstream exon and the substitution of specific sequences affect pre-mRNA splicing in vitro. Mol Cell Biol. 1988 Feb;8(2):860–866. doi: 10.1128/mcb.8.2.860. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Graham F. L., van der Eb A. J. A new technique for the assay of infectivity of human adenovirus 5 DNA. Virology. 1973 Apr;52(2):456–467. doi: 10.1016/0042-6822(73)90341-3. [DOI] [PubMed] [Google Scholar]
  12. Green M. R. Pre-mRNA splicing. Annu Rev Genet. 1986;20:671–708. doi: 10.1146/annurev.ge.20.120186.003323. [DOI] [PubMed] [Google Scholar]
  13. Hwang L. S., Park J., Gilboa E. Role of intron-contained sequences in formation of moloney murine leukemia virus env mRNA. Mol Cell Biol. 1984 Nov;4(11):2289–2297. doi: 10.1128/mcb.4.11.2289. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Katz R. A., Kotler M., Skalka A. M. cis-acting intron mutations that affect the efficiency of avian retroviral RNA splicing: implication for mechanisms of control. J Virol. 1988 Aug;62(8):2686–2695. doi: 10.1128/jvi.62.8.2686-2695.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Katz R. A., Skalka A. M. Control of retroviral RNA splicing through maintenance of suboptimal processing signals. Mol Cell Biol. 1990 Feb;10(2):696–704. doi: 10.1128/mcb.10.2.696. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Klinz F. J., Gallwitz D. Size and position of intervening sequences are critical for the splicing efficiency of pre-mRNA in the yeast Saccharomyces cerevisiae. Nucleic Acids Res. 1985 Jun 11;13(11):3791–3804. doi: 10.1093/nar/13.11.3791. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Lazo P. A., Prasad V., Tsichlis P. N. Splice acceptor site for the env message of Moloney murine leukemia virus. J Virol. 1987 Jun;61(6):2038–2041. doi: 10.1128/jvi.61.6.2038-2041.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Mann R., Baltimore D. Varying the position of a retrovirus packaging sequence results in the encapsidation of both unspliced and spliced RNAs. J Virol. 1985 May;54(2):401–407. doi: 10.1128/jvi.54.2.401-407.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Mann R., Mulligan R. C., Baltimore D. Construction of a retrovirus packaging mutant and its use to produce helper-free defective retrovirus. Cell. 1983 May;33(1):153–159. doi: 10.1016/0092-8674(83)90344-6. [DOI] [PubMed] [Google Scholar]
  20. Nash M. A., Brizzard B. L., Wong J. L., Murphy E. C., Jr Murine sarcoma virus ts110 RNA transcripts: origin from a single proviral DNA and sequence of the gag-mos junctions in both the precursor and spliced viral RNAs. J Virol. 1985 Feb;53(2):624–633. doi: 10.1128/jvi.53.2.624-633.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Nash M., Brown N. V., Wong J. L., Arlinghaus R. B., Murphy E. C., Jr S1 nuclease mapping of viral RNAs from a temperature-sensitive transformation mutant of murine sarcoma virus. J Virol. 1984 May;50(2):478–488. doi: 10.1128/jvi.50.2.478-488.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Padgett R. A., Grabowski P. J., Konarska M. M., Seiler S., Sharp P. A. Splicing of messenger RNA precursors. Annu Rev Biochem. 1986;55:1119–1150. doi: 10.1146/annurev.bi.55.070186.005351. [DOI] [PubMed] [Google Scholar]
  23. Reed R., Maniatis T. A role for exon sequences and splice-site proximity in splice-site selection. Cell. 1986 Aug 29;46(5):681–690. doi: 10.1016/0092-8674(86)90343-0. [DOI] [PubMed] [Google Scholar]
  24. Reed R., Maniatis T. The role of the mammalian branchpoint sequence in pre-mRNA splicing. Genes Dev. 1988 Oct;2(10):1268–1276. doi: 10.1101/gad.2.10.1268. [DOI] [PubMed] [Google Scholar]
  25. Reed R. The organization of 3' splice-site sequences in mammalian introns. Genes Dev. 1989 Dec;3(12B):2113–2123. doi: 10.1101/gad.3.12b.2113. [DOI] [PubMed] [Google Scholar]
  26. Schirm S., Moscovici G., Bishop J. M. A temperature-sensitive phenotype of avian myeloblastosis virus: determinants that influence the production of viral mRNAs. J Virol. 1990 Feb;64(2):767–773. doi: 10.1128/jvi.64.2.767-773.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Shinnick T. M., Lerner R. A., Sutcliffe J. G. Nucleotide sequence of Moloney murine leukaemia virus. Nature. 1981 Oct 15;293(5833):543–548. doi: 10.1038/293543a0. [DOI] [PubMed] [Google Scholar]
  28. Smith C. W., Nadal-Ginard B. Mutually exclusive splicing of alpha-tropomyosin exons enforced by an unusual lariat branch point location: implications for constitutive splicing. Cell. 1989 Mar 10;56(5):749–758. doi: 10.1016/0092-8674(89)90678-8. [DOI] [PubMed] [Google Scholar]
  29. Smith C. W., Porro E. B., Patton J. G., Nadal-Ginard B. Scanning from an independently specified branch point defines the 3' splice site of mammalian introns. Nature. 1989 Nov 16;342(6247):243–247. doi: 10.1038/342243a0. [DOI] [PubMed] [Google Scholar]
  30. Stoltzfus C. M., Fogarty S. J. Multiple regions in the Rous sarcoma virus src gene intron act in cis to affect the accumulation of unspliced RNA. J Virol. 1989 Apr;63(4):1669–1676. doi: 10.1128/jvi.63.4.1669-1676.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Thompson-Jäger S., Domdey H. Yeast pre-mRNA splicing requires a minimum distance between the 5' splice site and the internal branch acceptor site. Mol Cell Biol. 1987 Nov;7(11):4010–4016. doi: 10.1128/mcb.7.11.4010. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Tsurushita N., Korn L. J. Effects of intron length on differential processing of mouse mu heavy-chain mRNA. Mol Cell Biol. 1987 Jul;7(7):2602–2605. doi: 10.1128/mcb.7.7.2602. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Van Beveren C., van Straaten F., Galleshaw J. A., Verma I. M. Nucleotide sequence of the genome of a murine sarcoma virus. Cell. 1981 Nov;27(1 Pt 2):97–108. doi: 10.1016/0092-8674(81)90364-0. [DOI] [PubMed] [Google Scholar]
  34. Wieringa B., Hofer E., Weissmann C. A minimal intron length but no specific internal sequence is required for splicing the large rabbit beta-globin intron. Cell. 1984 Jul;37(3):915–925. doi: 10.1016/0092-8674(84)90426-4. [DOI] [PubMed] [Google Scholar]
  35. de Mars M., Cizdziel P. E., Murphy E. C., Jr Activation of thermosensitive RNA splicing and production of a heat-labile P85gag-mos kinase by the introduction of a specific deletion in murine sarcoma virus-124 DNA. J Virol. 1988 Jun;62(6):1907–1916. doi: 10.1128/jvi.62.6.1907-1916.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. van Santen V. L., Spritz R. A. mRNA precursor splicing in vivo: sequence requirements determined by deletion analysis of an intervening sequence. Proc Natl Acad Sci U S A. 1985 May;82(9):2885–2889. doi: 10.1073/pnas.82.9.2885. [DOI] [PMC free article] [PubMed] [Google Scholar]

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