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. 1995 Dec;69(12):7775–7781. doi: 10.1128/jvi.69.12.7775-7781.1995

Host-dependent evolution of the Sindbis virus promoter for subgenomic mRNA synthesis.

J M Hertz 1, H V Huang 1
PMCID: PMC189720  PMID: 7494288

Abstract

Alphaviruses are alternately transmitted between arthropod and vertebrate hosts. In each host, the virus transcribes a subgenomic mRNA that encodes the viral structural proteins which encapsidate the genome to form progeny virions. Transcription initiates at an internal site called the promoter. To determine if promoter utilization varies in mammalian versus mosquito cells, we used these cells as hosts to select for active promoters among a library of different mutant promoters. Compared with that in BHK-21 cells, selection was more rapid in mosquito (C7-10) cells, with much less diversity of promoters remaining after fewer passages. Thus, promoter selection is host dependent. With further passaging, both BHK-21 and C7-10 cells selected for similar sequences that closely resemble the wild-type promoter sequence. The difference in the rates of selection is not because BHK-21-derived promoters cannot function in mosquito cells. Instead, part of the host dependence is probably due to posttranscriptional differences between BHK-21 and C7-10 cells that may require more active promoters in mosquito cells. Part of the host dependence may also be attributed to the decreased rate of transcription versus that of replication in mosquito cells. This change in regulation of subgenomic to genomic RNA synthesis appears to correlate with the extent of cleavage or pausing of the genomic RNA synthesis at or close to the promoter.

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Selected References

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  1. Bonatti S., Migliaccio G., Simons K. Palmitylation of viral membrane glycoproteins takes place after exit from the endoplasmic reticulum. J Biol Chem. 1989 Jul 25;264(21):12590–12595. [PubMed] [Google Scholar]
  2. Condreay L. D., Brown D. T. Suppression of RNA synthesis by a specific antiviral activity in Sindbis virus-infected Aedes albopictus cells. J Virol. 1988 Jan;62(1):346–348. doi: 10.1128/jvi.62.1.346-348.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Durbin R. K., Stollar V. A mutant of sindbis virus with a host-dependent defect in maturation associated with hyperglycosylation of E2. Virology. 1984 Jun;135(2):331–344. doi: 10.1016/0042-6822(84)90190-9. [DOI] [PubMed] [Google Scholar]
  4. Durbin R., Kane A., Stollar V. A mutant of Sindbis virus with altered plaque morphology and a decreased ratio of 26 S:49 S RNA synthesis in mosquito cells. Virology. 1991 Jul;183(1):306–312. doi: 10.1016/0042-6822(91)90143-y. [DOI] [PubMed] [Google Scholar]
  5. Erwin C., Brown D. T. Requirement of cell nucleus for Sindbis virus replication in cultured Aedes albopictus cells. J Virol. 1983 Feb;45(2):792–799. doi: 10.1128/jvi.45.2.792-799.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Froshauer S., Kartenbeck J., Helenius A. Alphavirus RNA replicase is located on the cytoplasmic surface of endosomes and lysosomes. J Cell Biol. 1988 Dec;107(6 Pt 1):2075–2086. doi: 10.1083/jcb.107.6.2075. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Hahn Y. S., Grakoui A., Rice C. M., Strauss E. G., Strauss J. H. Mapping of RNA- temperature-sensitive mutants of Sindbis virus: complementation group F mutants have lesions in nsP4. J Virol. 1989 Mar;63(3):1194–1202. doi: 10.1128/jvi.63.3.1194-1202.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Hahn Y. S., Strauss E. G., Strauss J. H. Mapping of RNA- temperature-sensitive mutants of Sindbis virus: assignment of complementation groups A, B, and G to nonstructural proteins. J Virol. 1989 Jul;63(7):3142–3150. doi: 10.1128/jvi.63.7.3142-3150.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Hertz J. M., Huang H. V. Evolution of the Sindbis virus subgenomic mRNA promoter in cultured cells. J Virol. 1995 Dec;69(12):7768–7774. doi: 10.1128/jvi.69.12.7768-7774.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Hertz J. M., Huang H. V. Utilization of heterologous alphavirus junction sequences as promoters by Sindbis virus. J Virol. 1992 Feb;66(2):857–864. doi: 10.1128/jvi.66.2.857-864.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Igarashi A., Stollar V. Failure of defective interfering particles of Sindbis virus produced in BHK or chicken cells to affect viral replication in Aedes albopictus cells. J Virol. 1976 Aug;19(2):398–408. doi: 10.1128/jvi.19.2.398-408.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Keränen S., Käriäinen L. Functional defects of RNA-negative temperature-sensitive mutants of Sindbis and Semliki Forest viruses. J Virol. 1979 Oct;32(1):19–29. doi: 10.1128/jvi.32.1.19-29.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. LaStarza M. W., Lemm J. A., Rice C. M. Genetic analysis of the nsP3 region of Sindbis virus: evidence for roles in minus-strand and subgenomic RNA synthesis. J Virol. 1994 Sep;68(9):5781–5791. doi: 10.1128/jvi.68.9.5781-5791.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Lemm J. A., Rice C. M. Roles of nonstructural polyproteins and cleavage products in regulating Sindbis virus RNA replication and transcription. J Virol. 1993 Apr;67(4):1916–1926. doi: 10.1128/jvi.67.4.1916-1926.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Lemm J. A., Rümenapf T., Strauss E. G., Strauss J. H., Rice C. M. Polypeptide requirements for assembly of functional Sindbis virus replication complexes: a model for the temporal regulation of minus- and plus-strand RNA synthesis. EMBO J. 1994 Jun 15;13(12):2925–2934. doi: 10.1002/j.1460-2075.1994.tb06587.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Levis R., Schlesinger S., Huang H. V. Promoter for Sindbis virus RNA-dependent subgenomic RNA transcription. J Virol. 1990 Apr;64(4):1726–1733. doi: 10.1128/jvi.64.4.1726-1733.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Liljeström P., Lusa S., Huylebroeck D., Garoff H. In vitro mutagenesis of a full-length cDNA clone of Semliki Forest virus: the small 6,000-molecular-weight membrane protein modulates virus release. J Virol. 1991 Aug;65(8):4107–4113. doi: 10.1128/jvi.65.8.4107-4113.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Mi S., Durbin R., Huang H. V., Rice C. M., Stollar V. Association of the Sindbis virus RNA methyltransferase activity with the nonstructural protein nsP1. Virology. 1989 Jun;170(2):385–391. doi: 10.1016/0042-6822(89)90429-7. [DOI] [PubMed] [Google Scholar]
  19. Mi S., Stollar V. Both amino acid changes in nsP1 of Sindbis virusLM21 contribute to and are required for efficient expression of the mutant phenotype. Virology. 1990 Oct;178(2):429–434. doi: 10.1016/0042-6822(90)90340-w. [DOI] [PubMed] [Google Scholar]
  20. Mi S., Stollar V. Expression of Sindbis virus nsP1 and methyltransferase activity in Escherichia coli. Virology. 1991 Sep;184(1):423–427. doi: 10.1016/0042-6822(91)90862-6. [DOI] [PubMed] [Google Scholar]
  21. Miller M. L., Brown D. T. The distribution of Sindbis virus proteins in mosquito cells as determined by immunofluorescence and immunoelectron microscopy. J Gen Virol. 1993 Feb;74(Pt 2):293–298. doi: 10.1099/0022-1317-74-2-293. [DOI] [PubMed] [Google Scholar]
  22. Ou J. H., Rice C. M., Dalgarno L., Strauss E. G., Strauss J. H. Sequence studies of several alphavirus genomic RNAs in the region containing the start of the subgenomic RNA. Proc Natl Acad Sci U S A. 1982 Sep;79(17):5235–5239. doi: 10.1073/pnas.79.17.5235. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Presley J. F., Polo J. M., Johnston R. E., Brown D. T. Proteolytic processing of the Sindbis virus membrane protein precursor PE2 is nonessential for growth in vertebrate cells but is required for efficient growth in invertebrate cells. J Virol. 1991 Apr;65(4):1905–1909. doi: 10.1128/jvi.65.4.1905-1909.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Raju R., Huang H. V. Analysis of Sindbis virus promoter recognition in vivo, using novel vectors with two subgenomic mRNA promoters. J Virol. 1991 May;65(5):2501–2510. doi: 10.1128/jvi.65.5.2501-2510.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Rice C. M., Strauss J. H. Nucleotide sequence of the 26S mRNA of Sindbis virus and deduced sequence of the encoded virus structural proteins. Proc Natl Acad Sci U S A. 1981 Apr;78(4):2062–2066. doi: 10.1073/pnas.78.4.2062. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Sarver N., Stollar V. Sindbis virus-induced cytopathic effect in clones of Aedes albopictus (Singh) cells. Virology. 1977 Jul 15;80(2):390–400. doi: 10.1016/s0042-6822(77)80014-7. [DOI] [PubMed] [Google Scholar]
  27. Sawicki D. L., Kaariainen L., Lambek C., Gomatos P. J. Mechanism for control of synthesis of Semliki Forest virus 26S and 42s RNA. J Virol. 1978 Jan;25(1):19–27. doi: 10.1128/jvi.25.1.19-27.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Sawicki D. L., Sawicki S. G. Functional analysis of the A complementation group mutants of Sindbis HR virus. Virology. 1985 Jul 15;144(1):20–34. doi: 10.1016/0042-6822(85)90301-0. [DOI] [PubMed] [Google Scholar]
  29. Schärer C. G., Naim H. Y., Koblet H. Palmitoylation of Semliki Forest virus glycoproteins in insect cells (C6/36) occurs in an early compartment and is coupled to the cleavage of the precursor p62. Arch Virol. 1993;132(3-4):237–254. doi: 10.1007/BF01309536. [DOI] [PubMed] [Google Scholar]
  30. Simmons D. T., Strauss J. H. Replication of Sindbis virus. II. Multiple forms of double-stranded RNA isolated from infected cells. J Mol Biol. 1972 Nov 28;71(3):615–631. doi: 10.1016/s0022-2836(72)80027-5. [DOI] [PubMed] [Google Scholar]
  31. Strauss J. H., Strauss E. G. The alphaviruses: gene expression, replication, and evolution. Microbiol Rev. 1994 Sep;58(3):491–562. doi: 10.1128/mr.58.3.491-562.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Ubol S., Levine B., Lee S. H., Greenspan N. S., Griffin D. E. Roles of immunoglobulin valency and the heavy-chain constant domain in antibody-mediated downregulation of Sindbis virus replication in persistently infected neurons. J Virol. 1995 Mar;69(3):1990–1993. doi: 10.1128/jvi.69.3.1990-1993.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Wang Y. F., Sawicki S. G., Sawicki D. L. Alphavirus nsP3 functions to form replication complexes transcribing negative-strand RNA. J Virol. 1994 Oct;68(10):6466–6475. doi: 10.1128/jvi.68.10.6466-6475.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]

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