1887

Journal of General Virology header logo

Volume 72, Issue 6

Regulated M1 mRNA splicing in influenza virus-infected cells Free

Abstract

Influenza virus RNA segment 7 generates three poly(A) RNAs, M1 mRNA, M2 mRNA and mRNA3, the last of which has almost no coding capacity; M2 mRNA and mRNA3 derive from M1 mRNA by removal of a single intron. The kinetics of M1 and M2 mRNA accumulation in the cytoplasm of productively infected cells were studied by means of a quantitative RNA protection assay; the ratio of M2 mRNA to M1 mRNA increased 2.7-fold during the course of infection. To analyse the basis for this change, the kinetics of M1 and M2 mRNA synthesis and nuclear accumulation, their stability and nucleocytoplasmic transport were studied. Under the experimental conditions used, the synthesis of segment 7-specific RNA showed a peak at 4 h post-infection and continued later at a slower rate. The half-lives of M1 and M2 mRNAs were indistinguishable (2.73 h for M1 mRNA and 2.70 h for M2 mRNA) and the kinetics of nucleocytoplasmic transport or showed no preference for either mRNA early or late in infection. Consequently, regulation at the level of mRNA splicing is proposed. Using the mRNA synthesis and stability data, a simulation was performed to predict the change in splicing efficiency required to account for the mRNA accumulation results. The best fit was obtained when splicing efficiency changed about 20 times during a period in which viral gene expression was maximal.

  • Received:
  • Accepted:
  • Published Online:
© Journal of General Virology 1991
Loading

Article metrics loading...

/content/journal/jgv/10.1099/0022-1317-72-6-1301
1991-06-01
2024-04-19
Loading full text...

Full text loading...

/deliver/fulltext/jgv/72/6/JV0720061301.html?itemId=/content/journal/jgv/10.1099/0022-1317-72-6-1301&mimeType=html&fmt=ahah

References

  1. Agris C. H., Nemeroff M. E., Krug R. M. 1989; A block in mammalian splicing occurring after formation of large complexes containing Ul, U2, U4, U5 and U6 small nuclear ribonucleopro-teins. Molecular and Cellular Biology 9:259–267
     [Google Scholar]
  2. Allen H., McCauley J., Waterfield M., Gething M. J. 1980; Influenza virus RNA segment 7 has the coding capacity for two polypeptides. Virology 107:548–551
     [Google Scholar]
  3. Baxer B. S. 1989; Sex in flies: the splice of life. Nature, London 340:521–524
     [Google Scholar]
  4. Bradshaw G. L., Schwartz C. D., Schlesinger R. W. 1990; Replication of H1N1 influenza virus in cultured mouse embryo brain cells: virus strain and cell differentiation affect synthesis of proteins encoded in RNA segments 7 and 8 and efficiency of mRN A splicing. Virology 176:390–402
     [Google Scholar]
  5. Breitbart R. E., Andreadis A., Nadal-Girard B. 1987; Alternative splicing: a ubiquitous mechanism for the generation of multiple protein isoforms from single genes. Annual Review of Biochemistry 56:467–495
     [Google Scholar]
  6. Chang D. D., Sharp P. A. 1989; Regulation by HIV rev depends upon recognition of splice sites. Cell 59:789–795
     [Google Scholar]
  7. Cullen B. R., Greene W. C. 1989; Regulatory pathways governing HIV-1 replication. Cell 58:423–426
     [Google Scholar]
  8. Dayton E. T., Powell D. M., Dayton A. I. 1989; Functional analysis of CAR, the target sequence for the REV protein of HIV-1. Science 246:1625–1629
     [Google Scholar]
  9. Emerman M., Vazeux R., Peden K. 1989; The rev gene product of human immunodeficiency virus affects envelope-specific RNA localization. Cell 57:1155–1165
     [Google Scholar]
  10. Enami M., Fukuda R., Ishihama A. 1985; Transcription and replication of eight RNA segments of influenza virus. Virology 142:68–77
     [Google Scholar]
  11. Feinberg M. B., Jarret R. F., Aldovini A., Gallo R. C., Wong-Staal F. 1986; HTLV-III expression and production involve complex regulation at the levels of splicing and translation of viral RNA. Cell 46:807–817
     [Google Scholar]
  12. Goux-Pelletan M., Libri D., Aubenton-Carafa Y., Fiszman M., Brody E., Marie J. 1990; In vitro splicing of mutually exclusive exons from the chicken β-tropomysin gene: role of the branch point location and very long pyrimidine stretch. EMBO Journal 9:241–249
     [Google Scholar]
  13. Green M. R. 1986; Pre-mRNA splicing. Annual Review of Genetics 20:671–708
     [Google Scholar]
  14. Green M. R., Zapp M. L. 1989; Revving up gene expression. Nature, London 338:200–201
     [Google Scholar]
  15. Gregoriades A., Frangione B. 1981; Insertion of influenza M protein into the viral lipid bilayer and localization of the site of insertion. Journal of Virology 40:323–328
     [Google Scholar]
  16. Hanahan D. 1985; Techniques for transformation of E. coli . In DNA Cloning: A Practical Approach vol 1 pp 109–136 Edited by Glover D. M. Oxford: IRL Press;
     [Google Scholar]
  17. Hay A. J., Wolstenholme A. J., Skehel J. J., Smith M. H. 1985; The molecular basis of the specific anti-influenza action of amantadine. EMBO Journal 4:3021–3024
     [Google Scholar]
  18. Hodgkin J. 1989; Drosophila sex determination: a cascade of regulated splicing. Cell 56:905–906
     [Google Scholar]
  19. Inglis S. C., Brown C. M. 1981; Spliced and unspliced RNAs encoded by virion RNA segment 7 of influenza virus. Nucleic Acids Research 9:2727–2740
     [Google Scholar]
  20. Inglis S. C., Brown C. M. 1984; Differences in the control of virus mRNA splicing during permissive or abortive infection with influenza A (fowl plague) virus. Journal of General Virology 65:153164
     [Google Scholar]
  21. Katz R. A., Skalka A. M. 1990; Control of retroviral RNA splicing through maintenance of suboptimal processing signals. Molecular and Cellular Biology 10:696–704
     [Google Scholar]
  22. Katz R. A., Kotler M., Skalka A. M. 1988; Cis-acting intron mutations that affect the efficiency of avian retroviral RNA splicing: implication for mechanisms of control. Journal of Virology 62:2686–2695
     [Google Scholar]
  23. Lamb R. A. 1983; The influenza virus RNA segments and their encoded proteins. In Genetics of Influenza Viruses pp 27–69 Edited by Palese P., Kingsbury D. W. Wien: Springer-Verlag;
     [Google Scholar]
  24. Lamb R. A., Lai C. J. 1980; Sequence of interrupted and uninterrupted mRNAs and cloned DNA coding for the two overlapping nonstructural proteins of influenza virus. Cell 21:475485
     [Google Scholar]
  25. Lamb R. A., Lai C. J. 1982; Spliced and unspliced messenger RNAs synthesized from cloned influenza virus M DNA in an SV40 vector; expression of the influenza virus membrane protein (M1). Virology 123:237–256
     [Google Scholar]
  26. Lamb R. A., Lai C. J., Choppin P. W. 1981; Sequences of mRNAs derived from genome RNA segment 7 of influenza virus: colinear and interrupted mRNAs code for overlapping proteins. Proceedings of the National Academy of Sciences, U.S.A 78:4170–4174
     [Google Scholar]
  27. Lamb R. A., Zebedee S. L., Richardson C. D. 1985; Influenza virus M2 protein is an integral membrane protein expressed on the infected-cell surface. Cell 40:627–633
     [Google Scholar]
  28. Malim M. H., Hauber J., Le Sh.-Y., Maizel J. V., Cullen B. R. 1989; The HIV-1 rev trans-activator acts through a structural target sequence to activate nuclear export of unspliced viral mRNAs. Nature, London 338:254–257
     [Google Scholar]
  29. Maniatis T., Fritsch E. F., Sambrook J. 1982 Molecular Cloning: A Laboratory Manual New York: Cold Spring Harbor Laboratory;
     [Google Scholar]
  30. Melton P. A., Krieg P. A., Rebagliati M. R., Maniatis T., Zinn K., Green M. R. 1984; Efficient in vitro synthesis of biologically active RNA and RNA hybridization probes from plasmids containing a bacteriophage SP6 promoter. Nucleic Acids Research 12:7035–7056
     [Google Scholar]
  31. Mount S. M. 1982; A catalogue of splice junction sequences. Nucleic Acids Research 10:459–472
     [Google Scholar]
  32. Nagoshi R. N., Baker B. S. 1990; Regulation of sex-specific RNA splicing at the Drosophila doublesex gene: cis-acting mutations in exon sequences alter sex-specific RNA splicing patterns. Genes and Development 4:89–97
     [Google Scholar]
  33. Nagoshi R. N., McKeown M., Burtis K. C., Belote J. M., Baker B. S. 1988; The control of alternative splicing at genes regulating sexual differentiation in D. melanogaster . Cell 53:229–236
     [Google Scholar]
  34. Ortín J., Doerfler W. 1975; Transcription of the genome of adenovirus type 12. I. Viral mRNA in abortively infected and transformed ceils. Journal of Virology 15:27–35
     [Google Scholar]
  35. Ortín J., Najera R., Lopez C., Davila M., Domingo E. 1980; Genetic variability of Hong Kong (H3N2) influenza viruses; spontaneous mutations and their location in the viral genome. Gene 11:319–331
     [Google Scholar]
  36. Ortín J., Martínez C., Del Río L., Dávila M., López-Galíndez C., Villanueva N., Domingo E. 1983; Evolution of the nucleotide sequence of influenza virus RNA segment 7 during drift of the H3N2 subtype. Gene 23:233–239
     [Google Scholar]
  37. Plotch S. J., Krug R. M. 1986; In vitro splicing of influenza viral NS1 mRNA and NSl-β-globin chimeras: possible mechanisms for the control of viral mRNA splicing. Proceedings of the National Academy of Sciences, U.S.A 83:5444–5448
     [Google Scholar]
  38. Portela A. 1986 Clonage de genes del virus de la gripe en vectores que permiten su expresion en células de mamifero Ph.D. thesis, Universi-dad Autonoma, Madrid;
     [Google Scholar]
  39. Portela A., Melero J. A., Martínez C., Domingo E., Ortín J. 1985; Oriented synthesis and cloning of influenza virus nucleopro-tein cDNA that leads to its expression in mammalian cells. Virus Research 4:69–82
     [Google Scholar]
  40. Scholtissek C., Rott R. 1970; Synthesis in vivo of influenza virus plus and minus strand RNA and its preferential inhibition by antibiotics. Virology 40:989–996
     [Google Scholar]
  41. Schroeder H. C., Bachman M., Mueller W. E. G. 1989 Methods for Investigating Nucleo-Cytoplasmic Transport of RNA Stuttgart & New York: Gustav Fisher Verlag;
     [Google Scholar]
  42. Shapiro L. G., Gurney T. Jr, Krug R. M. 1987; Influenza virus gene expression: control mechanisms at early and late times of infection and nuclear-cytoplasmic transport of virus specific RNAs. Journal of Virology 61:764–773
     [Google Scholar]
  43. Smith C. W. J., Nadal-Ginard B. 1989; Mutually exclusive splicing of a-tropomyosin exons enforced by an unusual lariat branch point location: implications for constitutive splicing. Cell 56:749–758
     [Google Scholar]
  44. Smith D. B., Inglis S. C. 1985; Regulated production of an influenza virus spliced mRNA mediated by virus-specific products. EMBO Journal 4:2313–2319
     [Google Scholar]
  45. Sugrue R. J., Belshe R. B., Hay A. J. 1990a; Palmitoylation of the influenza A virus M2 protein. Virology 179:51–56
     [Google Scholar]
  46. Sugrue R. J., Bahadur G., Zambon M. C., Hall-Smith M., Douglas A. R., Hay A. J. 1990b; Specific alteration of the influenza haemagglutinin by amantadine. EMBO Journal 9:34693476
     [Google Scholar]
  47. Varmus H. 1988; Regulation of HIV and HTLV gene expression. Genes and Development 2:1055–1062
     [Google Scholar]
  48. Vieira J., Messing J. 1982; The pUC plasmids, an M13mp7-derived system for insertion mutagenesis and sequencing with universal primers. Gene 19:259–268
     [Google Scholar]
  49. Ye Z., Pal R., Fox J. W., Wagner R. R. 1987; Functional and antigenic domains of the matrix (M1) protein of influenza A virus. Journal of Virology 61:239–246
     [Google Scholar]
  50. Ye Z., Baylor N. W., Wagner R. R. 1989; Transcription-inhibition and RNA-binding domains of influenza A virus matrix protein mapped with anti-idiotypic antibodies and synthetic peptides. Journal of Virology 63:3586–3594
     [Google Scholar]
  51. Zebedee S. L., Lamb R. A. 1988; Influenza A virus M2 protein: monoclonal antibody restriction of virus growth and detection of M2 in the virions. Journal of Virology 62:2762–2772
     [Google Scholar]
  52. Zebedee S. L., Lamb R. A. 1989; Growth restriction of influenza A virus by M2 protein antibody is genetically linked to the M1 protein. Proceedings of the National Academy of Sciences, U.S.A 86:1061–1065
     [Google Scholar]
  53. Zebedee S. L., Richardson C. D., Lamb R. A. 1985; Characterization of the influenza virus M2 integral membrane protein and expression at the infected-cell surface from cloned cDNA. Journal of Virology 56:502–511
     [Google Scholar]
  54. Zvonarjev A. Y., Ghendon Y. Z. 1980; Influence of membrane (M) protein on influenza A virus virion transcriptase activity and its susceptibility to rimantadine. Journal of Virology 33:583–586
     [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jgv/10.1099/0022-1317-72-6-1301
Loading
Regulated M1 mRNA splicing in influenza virus-infected cells
J. Gen. Virol. 72, 1301 (1991); https://doi.org/10.1099/0022-1317-72-6-1301
/content/journal/jgv/10.1099/0022-1317-72-6-1301
/content/journal/jgv/10.1099/0022-1317-72-6-1301
Loading

Data & Media loading...

Most cited Most Cited RSS feed

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error