Abstract
In the accompanying report, we describe an in vitro polymerase assay based on reconstituted Thogoto virus (THOV) cores which provided evidence of a double-stranded vRNA promoter consisting of both the 3' and 5' sequences of vRNA (M. B. Leahy, J. T. Dessens, and P. A. Nuttall, J. Virol. 71:8347-8351, 1997). This system was used to investigate further the THOV vRNA promoter structure by using short, synthetic vRNA promoters. The results obtained show that interstrand base pairing between residues 10 and 11 of the 3' promoter arm with residues 11 and 12 of the 5' promoter arm, respectively, is important for promoter activity. In addition, intrastrand base pairing between residues 2 and 3 with residues 9 and 8 of the 5' promoter arm, respectively, was shown to be involved in promoter activity, while no evidence of intrastrand base pairing between residues 2 and 9 of the 3' promoter arm was obtained. These observations are consistent with a hook-like structure in the 5' promoter arm of the THOV promoter. The THOV cores were able to transcribe an influenza A virus (FLUA) vRNA-like promoter, as well as hybrid THOV-FLUA promoters. Hence, the THOV and FLUA vRNA promoters appear to be both structurally and functionally similar.
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Selected References
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- Albo C., Martín J., Portela A. The 5' ends of Thogoto virus (Orthomyxoviridae) mRNAs are homogeneous in both length and sequence. J Virol. 1996 Dec;70(12):9013–9017. doi: 10.1128/jvi.70.12.9013-9017.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Clerx J. P., Fuller F., Bishop D. H. Tick-borne viruses structurally similar to Orthomyxoviruses. Virology. 1983 May;127(1):205–219. doi: 10.1016/0042-6822(83)90384-7. [DOI] [PubMed] [Google Scholar]
- Flick R., Neumann G., Hoffmann E., Neumeier E., Hobom G. Promoter elements in the influenza vRNA terminal structure. RNA. 1996 Oct;2(10):1046–1057. [PMC free article] [PubMed] [Google Scholar]
- Fodor E., Pritlove D. C., Brownlee G. G. Characterization of the RNA-fork model of virion RNA in the initiation of transcription in influenza A virus. J Virol. 1995 Jul;69(7):4012–4019. doi: 10.1128/jvi.69.7.4012-4019.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Fodor E., Pritlove D. C., Brownlee G. G. The influenza virus panhandle is involved in the initiation of transcription. J Virol. 1994 Jun;68(6):4092–4096. doi: 10.1128/jvi.68.6.4092-4096.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Kim H. J., Fodor E., Brownlee G. G., Seong B. L. Mutational analysis of the RNA-fork model of the influenza A virus vRNA promoter in vivo. J Gen Virol. 1997 Feb;78(Pt 2):353–357. doi: 10.1099/0022-1317-78-2-353. [DOI] [PubMed] [Google Scholar]
- Leahy M. B., Dessens J. T., Nuttall P. A. In vitro polymerase activity of Thogoto virus: evidence for a unique cap-snatching mechanism in a tick-borne orthomyxovirus. J Virol. 1997 Nov;71(11):8347–8351. doi: 10.1128/jvi.71.11.8347-8351.1997. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Leahy M. B., Dessens J. T., Weber F., Kochs G., Nuttall P. A. The fourth genus in the Orthomyxoviridae: sequence analyses of two Thogoto virus polymerase proteins and comparison with influenza viruses. Virus Res. 1997 Aug;50(2):215–224. doi: 10.1016/s0168-1702(97)00072-5. [DOI] [PubMed] [Google Scholar]
- Morse M. A., Marriott A. C., Nuttall P. A. The glycoprotein of Thogoto virus (a tick-borne orthomyxo-like virus) is related to the baculovirus glycoprotein GP64. Virology. 1992 Feb;186(2):640–646. doi: 10.1016/0042-6822(92)90030-s. [DOI] [PubMed] [Google Scholar]
- Neumann G., Hobom G. Mutational analysis of influenza virus promoter elements in vivo. J Gen Virol. 1995 Jul;76(Pt 7):1709–1717. doi: 10.1099/0022-1317-76-7-1709. [DOI] [PubMed] [Google Scholar]
- Pringle C. R. Virus taxonomy 1996 - a bulletin from the Xth International Congress of Virology in Jerusalem. Arch Virol. 1996;141(11):2251–2256. doi: 10.1007/BF01718231. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Staunton D., Nuttall P. A., Bishop D. H. Sequence analyses of Thogoto viral RNA segment 3: evidence for a distant relationship between an arbovirus and members of the Orthomyxoviridae. J Gen Virol. 1989 Oct;70(Pt 10):2811–2817. doi: 10.1099/0022-1317-70-10-2811. [DOI] [PubMed] [Google Scholar]
- Weber F., Haller O., Kochs G. Nucleoprotein viral RNA and mRNA of Thogoto virus: a novel "cap-stealing" mechanism in tick-borne orthomyxoviruses? J Virol. 1996 Dec;70(12):8361–8367. doi: 10.1128/jvi.70.12.8361-8367.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Zheng H., Palese P., García-Sastre A. Nonconserved nucleotides at the 3' and 5' ends of an influenza A virus RNA play an important role in viral RNA replication. Virology. 1996 Mar 1;217(1):242–251. doi: 10.1006/viro.1996.0111. [DOI] [PubMed] [Google Scholar]