1887

Abstract

During the replication of Lelystad virus in alveolar lung macrophages, a 3′-coterminal nested set of six subgenomic RNAs (RNA2 to RNA7) is formed. These contain a common leader sequence derived from the 5′ non-coding region of the genomic RNA. In this study, the sequence of the junction sites, i.e. the sites where the leader sequence joins to the body of the subgenomic RNA, was determined for all six subgenomic RNAs. For each subgenomic RNA, six to nine cDNA clones were isolated by means of reverse transcription and PCR. The nucleotide sequence at the junction site was identical for all eight cDNA clones derived from subgenomic RNA4. However, heterogeneity was observed in the nucleotide sequence surrounding the junction sites of the cDNA clones derived from subgenomic RNAs 2, 3, 5, 6 and 7. This heterogeneity suggests that the fusion of the leader to the body of the subgenomic RNA may be imprecise. The junction sites of the six subgenomic RNAs had a conserved sequence motif of six nucleotides (UCAACC or a highly similar sequence). The distance between the junction site and the translation initiation codon of the downstream open reading frame varied from nine to 83 nucleotides.

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1993-08-01
2024-04-18
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References

  1. Baker SC, Lai MMC. 1990; Anin vitro system for the leader-primed transcription of the coronavirus mRNAs.. EMBO Journal 9:4173–4179
    [Google Scholar]
  2. Baric RS, Stohlman SA, Lai MMC. 1983; Characterization of replicative intermediate RNA of mouse hepatitis virus: presence of leader RNA sequences on nascent chains.. Journal of Virology 48:633–640
    [Google Scholar]
  3. Baric RS, Stohlman SA, Razavi MK, Lai MMC. 1985; Characterization of leader-related small RNAs in coronavirus-infected cells: further evidence for leader-primed mechanism of transcription.. Virus Research 3:19–33
    [Google Scholar]
  4. Cech TR. 1990; Self-splicing of group I introns.. Annual Review of Biochemistry 59:543–568
    [Google Scholar]
  5. Chen Z, Kuo L, Rowland RRR, Even C, Faaberg KS, Plagemann PGW. 1993; Sequences of 3′ end of genome and of 5′ end of open reading frame la of lactate dehydrogenase-elevating virus and common junction motifs between 5′ leader and bodies of seven subgenomic mRNAs.. Journal of General Virology 74:643–660
    [Google Scholar]
  6. de Vries AAF, Chirnside ED, Bredenbeek PJ, Gravestein LA, Horzinek MC, Spaan WJM. 1990; All subgenomic mRNAs of equine arteritis virus contain a common leader sequence.. Nucleic Acids Research 18:3241–3247
    [Google Scholar]
  7. Kuo L, Chen Z, Rowland RRR, Faaberg KS, Plagemann PGW. 1992; Lactate dehydrogenase-elevating virus (LDV): subgenomic mRNAs, mRNA leader and comparison of 3′ terminal sequences of two LDV isolates.. Virus Research 23:55–72
    [Google Scholar]
  8. Makino S, Stohlman SA, Lai MMC. 1986; Leader sequences of murine coronavirus mRNAs can be freely reassorted: evidence for the role of free leader RNA in transcription. . Proceedings of the National Academy of Sciences, U.S.A. 83:4202–4208
    [Google Scholar]
  9. Makino S, Shieh C, Lai MMC. 1988; Discontinuous transcription generates heterogeneity at the leader fusion sites of coronavirus mRNAs.. Journal of Virology 62:3870–3873
    [Google Scholar]
  10. Makino S, Joo M, Makino JK. 1991; A system for study of coronavirus mRNA synthesis: a regulated expressed subgenomic defective interfering RNA results from intergenic site insertion.. Journal of Virology 65:6031–6041
    [Google Scholar]
  11. Meulenberg JJM, FIulst MM, de Meder E, Moonen PLJM, den Besten A, de Kluyver EP, Wensvoort G, Moormann RJM. 1993; Lelystad virus, the causative agent of porcine epidemic abortion and respiratory syndrome (PEARS), is related to LDV and EAV.. Virology 192:62–72
    [Google Scholar]
  12. Sambrook J, Fritsch EF, Maniatis T. 1989 Molecular Cloning: A Laboratory Manual, 2nd edn.. New York: Cold Spring Harbor Laboratory.;
    [Google Scholar]
  13. Sawicki SG, Sawicki DL. 1990; Coronavirus transcription: subgenomic mouse hepatitis virus replicative intermediates function in RNA synthesis.. Journal of Virology 64:1050–1056
    [Google Scholar]
  14. Sethna PB, Hung SL, Brian DA. 1989; Coronavirus subgenomic minus-strand RNAs and the potential for mRNA replicons. . Proceedings of the National Academy of Sciences, U.S.A. 86:5626–5630
    [Google Scholar]
  15. Spaan WJM, Cavanagh D, Horzinek MC. 1988; Coronaviruses: structure and genome expression.. Journal of General Virology 69:2939–2952
    [Google Scholar]
  16. van Berlo MF, Rottier PJM, Rottier PJM, van der Zeijst BAM. 1986; Intracellular equine arteritis virus (EAV)-specific RNAs contain common sequences.. Virology 152:492–496
    [Google Scholar]
  17. Wensvoort G, Terpstra C, Pol JMA, Ter Laak EA, Ter Laak EA, Bloemraad M, de Kluyver EP, Kragten C, van Buiten L, den Besten A, Wagenaar F, Broekhuijsen JM, Moonen PLJM, Zetstra T, de Boer EA, Tibben HJ, de Jong MF, van’t Veld P, Groenland GJR, van Gennep JA, Voets MTH, Verheijden JHM, Braamskamp J. 1991; Mystery swine disease in the Netherlands: the isolation of Lelystad vims.. Veterinary Quarterly 13:121–130
    [Google Scholar]
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