Abstract

Human respiratory syncytial (RS) virus, an enveloped virus that contains a single negative-sense strand of genomic RNA, is an important agent of pediatric respiratory tract disease. Previously, complete cDNAs and complete nucleotide sequences were obtained for nine of the ten known viral mRNAs. Here, synthetic oligodeoxynucleotides were used to direct dideoxynucleotide sequencing of intergenic and flanking regions in the viral genome. The results confirmed the sequences of the gene termini obtained from the cDNAs, showing that the nine viral mRNAs, initiate with the conserved sequence 5 prime GGGGCAAAUA.A.U.. and terminate with the conserved sequences 5 prime ... AGUA.UA(N)1-4-polyA. Comparison of the intergenic and flanking sequences with the complete mRNA sequences established unambiguously the 3' to 5' order of the nine genes on the viral genome. Each gene was immediately followed (in genome-sense) by an oligo U tract of 4-7 residues that might direct synthesis of poly A tails of the mRNAs by a reiterative copying mechanism. The intergenic regions varied in length from 1 to 52 nucleotides and displayed no obvious sequence conservation except that in all cases the last nucleotide (in genome-sense) was an A residue.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Intramural Research (Z01)
Project #
1Z01AI000368-03
Application #
4688502
Study Section
Project Start
Project End
Budget Start
Budget End
Support Year
3
Fiscal Year
1985
Total Cost
Indirect Cost

  Institution

Name
Niaid Extramural Activities
Department
Type
DUNS #
City
State
Country
United States
Zip Code

  Publications

Melendi, Guillermina A; Zavala, Fidel; Buchholz, Ursula J et al. (2007) Mapping and characterization of the primary and anamnestic H-2(d)-restricted cytotoxic T-lymphocyte response in mice against human metapneumovirus. J Virol 81:11461-7
Buchholz, Ursula J; Nagashima, Kunio; Murphy, Brian R et al. (2006) Live vaccines for human metapneumovirus designed by reverse genetics. Expert Rev Vaccines 5:695-706
Biacchesi, Stephane; Pham, Quynh N; Skiadopoulos, Mario H et al. (2006) Modification of the trypsin-dependent cleavage activation site of the human metapneumovirus fusion protein to be trypsin independent does not increase replication or spread in rodents or nonhuman primates. J Virol 80:5798-806
Biacchesi, Stephane; Skiadopoulos, Mario H; Yang, Lijuan et al. (2005) Rapid human metapneumovirus microneutralization assay based on green fluorescent protein expression. J Virol Methods 128:192-7
Buchholz, Ursula J; Biacchesi, Stephane; Pham, Quynh N et al. (2005) Deletion of M2 gene open reading frames 1 and 2 of human metapneumovirus: effects on RNA synthesis, attenuation, and immunogenicity. J Virol 79:6588-97
Pham, Quynh N; Biacchesi, Stephane; Skiadopoulos, Mario H et al. (2005) Chimeric recombinant human metapneumoviruses with the nucleoprotein or phosphoprotein open reading frame replaced by that of avian metapneumovirus exhibit improved growth in vitro and attenuation in vivo. J Virol 79:15114-22
Biacchesi, Stephane; Skiadopoulos, Mario H; Yang, Lijuan et al. (2004) Recombinant human Metapneumovirus lacking the small hydrophobic SH and/or attachment G glycoprotein: deletion of G yields a promising vaccine candidate. J Virol 78:12877-87
Biacchesi, Stephane; Skiadopoulos, Mario H; Tran, Kim C et al. (2004) Recovery of human metapneumovirus from cDNA: optimization of growth in vitro and expression of additional genes. Virology 321:247-59
Buchholz, Ursula J; Bukreyev, Alexander; Yang, Lijuan et al. (2004) Contributions of the structural proteins of severe acute respiratory syndrome coronavirus to protective immunity. Proc Natl Acad Sci U S A 101:9804-9
Biacchesi, Stephane; Skiadopoulos, Mario H; Boivin, Guy et al. (2003) Genetic diversity between human metapneumovirus subgroups. Virology 315:1-9

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