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Mutational analysis of the gene start sequences of pneumonia virus of mice

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Dibben, Oliver and Easton, A. J. (Andrew J.). (2007) Mutational analysis of the gene start sequences of pneumonia virus of mice. Virus Research, Vol.130 (No.1-2). pp. 303-309. ISSN 0168-1702

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Official URL: http://dx.doi.org/10.1016/j.virusres.2007.06.009

Abstract

The transcriptional start sequence of pneumonia virus of mice is more variable than that of the other pneumoviruses, with five different nine-base gene start (GS) sequences found in the PVM genome. The sequence requirements of the PVM gene start signal, and the efficiency of transcriptional initiation of the different virus genes, was investigated using a reverse genetics approach with a minigenome construct containing two reporter genes. A series of GS mutants were created, where each of the nine bases of the gene start consensus sequence of a reporter gene was changed to every other possible base, and the resulting effect on initiation of transcription was assayed. Nucleotide positions 1, 2 and 7 were found to be most sensitive to mutation whilst positions 4, 5 and 9 were relatively insensitive. The L gene GS sequence was found to have only 20% of the activity of the consensus sequence whilst the published M2 gene start sequence was found to be non-functional. A minigenome construct in which the two reporter genes were separated by the F-M2 gene junction of PVM was used to confirm the presence of two alternative, functional, GS sequences that could both drive the transcription of the PVM M2 gene.

Item Type:	Journal Article
Subjects:	Q Science > QR Microbiology > QR355 Virology
Divisions:	Faculty of Science > Life Sciences (2010- ) > Biological Sciences ( -2010)
Library of Congress Subject Headings (LCSH):	Viral pneumonia -- Genetic aspects, Mice -- Virus diseases -- Genetic aspects, Paramyxoviruses -- Genetics
Journal or Publication Title:	Virus Research
Publisher:	Elsevier Science BV
ISSN:	0168-1702
Official Date:	December 2007
Volume:	Vol.130
Number:	No.1-2
Number of Pages:	7
Page Range:	pp. 303-309
Identification Number:	10.1016/j.virusres.2007.06.009
Status:	Peer Reviewed
Publication Status:	Published
Access rights to Published version:	Open Access
Funder:	Biotechnology and Biological Sciences Research Council (Great Britain) (BBSRC)
References:	Ahmadian, G., Chambers, P. and Easton, A.J. (1999) Detection and characterisation of proteins encoded by the second ORF of the M2 gene of pneumoviruses. J. Gen. Virol. 80, 2011-2016. Barr, J., Chambers, P., Pringle, C.R. and Easton, A.J. (1991). Sequence of the major nucleocapsid protein gene of pneumonia virus of mice: sequence comparisons suggest structural homology between nucleocapsid proteins of pneumoviruses, paramyxoviruses, rhabdoviruses and filoviruses. J. Gen. Virol. 72, 677-685. Bonville, C.A., Easton, A.J., Rosenberg, H.F. and Domachowske, J.B. (2003) Altered pathogenesis of severe pneumovirus infection in response to combined antiviral and specific immunomodulatory agents. J Virol. 77(2), 1237-44. Buchholz, U.J., Finke, S. and Conzelmann, K.-K. (1999) Generation of bovine respiratory syncytial virus (BRSV) from cDNA: BRSV NS2 is not essential for virus replication in tissue culture, and the human RSV leader region acts as a functional BRSV genome promoter. J. Virol. 73, 251-259. Chambers, P., Barr, J., Pringle, C.R. and Easton, A.J. (1990a) Molecular cloning of pneumonia virus of mice. J. Virol.64, 1869-1872. Chambers, P., Matthews, D.A., Pringle, C.R. and Easton, A.J. (1990b) The nucleotide sequences of intergenic regions between nine genes of pneumonia virus of mice establish the physical order of these genes in the viral genome. Vir. Res. 18, 263-270. Collins, P.L., Dickens, L.E., Buckler-White, A., Olmsted, R.A., Spriggs, M.K., Camargo, E. and Coelingh, K.L.v.W. (1986) Nucleotide sequences from the gene junctions of human respiratory syncytial virus reveal distinctive features of intergenic structure and gene order. Proc.Natl. Acad. Sci. USA 83, 4594-8. Collins, P.L., Hill, M.G., Cristina, J. and Grosfeld, H. (1996) Transcription elongation factor of respiratory syncytial virus, a nonsegmented negative-strand RNA virus. Proc.Natl. Acad. Sci. USA 93, 81-85. Collins, P.L., Olmsted, R.A., Spriggs, M.K., Johnson, P.R. and Buckler-White, A.J. (1987) Gene overlap and site-specific attenuation of transcription of the viral polymerase L gene of human respiratory syncytial virus. Proc.Natl. Acad. Sci. USA 84, 5134-8. Cook, P.M., Eglin, R.P. and Easton, A.J. (1998) Pathogenesis of pneumovirus infections in mice : detection of pneumonia virus of mice and human respiratory syncytial virus mRNA in lungs of infected mice by in-situ hybridisation. J. Gen. Virol. 79, 2411-2417. Dickens, L.E., Collins, P.L. and Wertz, G.W. (1984) Transcriptional mapping of human respiratory syncytial virus. J. Virol. 52, 364-9. Domachowske, J.B., Bonville, C.A., Dyer, K.D., Easton, A.J. and Rosenberg, H.F. (2000a) Pulmonary eosinophilia and production of MIP-1a are prominent responses to infection with pneumonia virus of mice. Cell. Immunol. 200, 98-104. Domachowske, J.B., Bonville, C.A., Easton, A.J. and Rosenberg, H.F. (2002) Differential expression of proinflammatory cytokine genes in vivo in response to pathogenic and nonpathogenic pneumovirus infections. J Infect Dis 186(1), 8-14. Domachowske, J.B., Bonville, C.A., Gao, J., Murphy, P.M., Easton, A.J. and Rosenberg, H.R. (2000b) The chemokine macrophage-inflammatory protein-1a and its receptor CCR1 control pulmonary inflammation and antiviral host defense in paramyxovirus infection. J.Immunol. 165, 2677-2682. Edworthy, N.L. and Easton, A.J. (2005) Mutational analysis of the avian pneumovirus conserved transcriptional gene start sequence identifying critical residues. J. Gen. Virol. 86, 3343-7. Grosfeld, H., Hill, M.G. and Collins, P.L. (1995) RNA replication by respiratory syncytial virus (RSV) is directed by the N, P and L proteins; transcription also occurs under these conditions but requires RSV superinfection for efficient synthesis of full-length mRNA. J. Virol. 69, 5677-5686. Iverson, L.E. and Rose, T.K. (1981) Localized attenuation and discontinuous synthesis during vesicular stomatitis virus transcription. Cell 23, 477-84. Kuo, L., Fearns, R. and Collins, P.L. (1997) Analysis of the gene start and gene end signals of human respiratory syncytial virus: quasi-templated initiation at position 1 of the encoded mRNA. J. Virol. 71, 4944-4953. Kuo, L., Grosfeld, H., Cristina, J., Hill, M.G. and Collins, P.L. (1996) Effect of mutations in the gene-start and gene-end sequence motifs on transcription of monocistronic and dicistronic minigenomes of respiratory syncytial virus. J. Virol. 70(10), 6892-6901. Li, J., Ling, R., Randhawa, J.S., Shaw, K., Davis, P.J., Juhasz, K., Pringle, C.R., Easton, A.J. and Cavanagh, D. (1996) Sequence of the nucleocapsid protein gene of subgroup-A and subgroup-B avian pneumoviruses. Vir. Res. 41(2), 185-191. Ling, R., Davis, P.J., Yu, Q.Z., Wood, C.M., Pringle, C.R., Cavanagh, D. and Easton, A.J. (1995) Sequence and in-vitro expression of the phosphoprotein gene of avian pneumovirus. Vir. Res. 36, 2-3. Ling, R., Easton, A.J. and Pringle, C.R. (1992) Sequence analysis of the 22K, SH and G genes of turkey rhinotracheitis virus and their intergenic regions reveals a gene order different from that of other pneumoviruses. J. Gen.Virol. 73, 1709-15. Marriott, A.C., Smith, J.M. and Easton, A.J. (2001) Fidelity of leader and trailer sequence usage by the respiratory syncytial virus and avian pneumovirus replication complexes. J. Virol. 75, 6265-6272. Marriott, A.C., Wilson, S.D., Randhawa, J.S. and Easton, A.J. (1999) A single amino acid substitution in the phosphoprotein of respiratory syncytial virus confers thermosensitivity in a reconstituted RNA polymerase system. J. Virol. 73, 5162-5165. Pringle, C.R. and Easton, A.J. (1997) Monopartite negative strand RNA genomes. Seminars in Virology 8, 49-57. Randhawa, J.S., Pringle, C.R. and Easton, A.J. (1996) Nucleotide-sequence of the matrix protein gene of a subgroup-B avian pneumovirus. Vir. Genes 12(2), 179-183. Thorpe, L.C. and Easton, A.J. (2005) Genome sequence of the non-pathogenic strain 15 of pneumonia virus of mice and comparison with the genome of the pathogenic strain J3666. J. Gen. Virol. 86, 159-169. Yu, Q., Davis, P.J., Barrett, T., Binns, M.M., Boursnell, M.E.G. and Cavanagh, D. (1991) Deduced amino acid sequence of the fusion glycoprotein of turkey rhinotracheitis virus has greater identity with that of human respiratory syncytial virus, a pneumovirus, than that of paramyxoviruses and morbilliviruses. Journal of General Virology 72, 75-81. Yu, Q., Davis, P.J., Brown, T.D.K. and Cavanagh, D. (1992) Sequence and in vitro expression of the M2 gene of turkey rhinotracheitis pneumovirus. J.Gen. Virol. 73, 1355-63.
URI:	http://wrap.warwick.ac.uk/id/eprint/30866