Identification of a Truncated Nucleoprotein in Avian Metapneumovirus-infected Cells Encoded by a Second AUG, In-frame to the Full-length Gene

Overview

Journal Virol J

Publisher Biomed Central

Specialty Microbiology

Date 2005 Apr 14

PMID 15826312

Citations 3

Authors

Rene Alvarez

Bruce S Seal

Affiliations

Soon will be listed here.

Abstract

Background: Avian metapneumoviruses (aMPV) cause an upper respiratory disease with low mortality, but high morbidity primarily in commercial turkeys. There are three types of aMPV (A, B, C) of which the C type is found only in the United States. Viruses related to aMPV include human, bovine, ovine, and caprine respiratory syncytial viruses and pneumonia virus of mice, as well as the recently identified human metapneumovirus (hMPV). The aMPV and hMPV have become the type viruses of a new genus within the Metapneumovirus. The aMPV nucleoprotein (N) amino acid sequences of serotypes A, B, and C were aligned for comparative analysis. Based on predicted antigenicity of consensus protein sequences, five aMPV-specific N peptides were synthesized for development of peptide-antigens and antisera.

Results: The presence of two aMPV nucleoprotein (N) gene encoded polypeptides was detected in aMPV/C/US/Co and aMPV/A/UK/3b infected Vero cells. Nucleoprotein 1 (N1) encoded from the first open reading frame (ORF) was predicted to be 394 amino acids in length for aMPV/C/US/Co and 391 amino acids in length for aMPV/A/UK/3b with approximate molecular weights of 43.3 kilodaltons and 42.7 kilodaltons, respectively. Nucleoprotein 2 (N2) was hypothesized to be encoded by a second downstream ORF in-frame with ORF1 and encoded a protein predicted to contain 328 amino acids for aMPV/C/US/Co or 259 amino acids for aMPV/A/UK/3b with approximate molecular weights of 36 kilodaltons and 28.3 kilodaltons, respectively. Peptide antibodies to the N-terminal and C-terminal portions of the aMPV N protein confirmed presence of these products in both aMPV/C/US/Co- and aMPV/A/UK/3b-infected Vero cells. N1 and N2 for aMPV/C/US/Co ORFs were molecularly cloned and expressed in Vero cells utilizing eukaryotic expression vectors to confirm identity of the aMPV encoded proteins.

Conclusion: This is the first reported identification of potential, accessory in-frame N2 ORF gene products among members of the Paramyxoviridae. Genomic sequence analyses of related members of the Pneumovirinae other than aMPV, including human respiratory syncytial virus and bovine respiratory syncytial virus demonstrated the presence of this second potential ORF among these agents.

Citing Articles

First report of type B in Iraqi broiler flocks with swollen head syndrome.

Al-Hasan B, Alhatami A, Abdulwahab H, Bustani G, Hameed M, Haider Jawad A Vet World. 2022; 15(1):16-21.

PMID: 35369601 PMC: 8924383. DOI: 10.14202/vetworld.2022.16-21.

Proteome Analysis in a Mammalian Cell line Reveals that PLK2 is Involved in Avian Metapneumovirus Type C (aMPV/C)-Induced Apoptosis.

Quan R, Wei L, Hou L, Wang J, Zhu S, Li Z Viruses. 2020; 12(4).

PMID: 32231136 PMC: 7232392. DOI: 10.3390/v12040375.

Deduced amino acid sequences surrounding the fusion glycoprotein cleavage site and of the carboxyl-terminus of haemagglutinin-neuraminidase protein of the avirulent thermostable vaccine strain I-2 of Newcastle disease virus.

Wambura P, Meers J, Kattenbelt J, Gould A, Spradbrow P Vet Res Commun. 2007; 31(1):105-12.

PMID: 17195928 DOI: 10.1007/s11259-006-3290-8.

References

Alvarez R, Jones L, Seal B, Kapczynski D, Tripp R . Serological cross-reactivity of members of the Metapneumovirus genus. Virus Res. 2004; 105(1):67-73. DOI: 10.1016/j.virusres.2004.04.019. View

Goyal S, Lauer D, Friendshuh K, Halvorson D . Seroprevalence of avian pneumovirus in Minnesota turkeys. Avian Dis. 2003; 47(3):700-6. DOI: 10.1637/6037. View

Seal B . Matrix protein gene nucleotide and predicted amino acid sequence demonstrate that the first US avian pneumovirus isolate is distinct from European strains. Virus Res. 1999; 58(1-2):45-52. DOI: 10.1016/s0168-1702(98)00098-7. View

Kozak M . Initiation of translation in prokaryotes and eukaryotes. Gene. 1999; 234(2):187-208. DOI: 10.1016/s0378-1119(99)00210-3. View

Inglis S, Brown C . Spliced and unspliced RNAs encoded by virion RNA segment 7 of influenza virus. Nucleic Acids Res. 1981; 9(12):2727-40. PMC: 326888. DOI: 10.1093/nar/9.12.2727. View

van den Hoogen B, de Jong J, Groen J, Kuiken T, de Groot R, Fouchier R . A newly discovered human pneumovirus isolated from young children with respiratory tract disease. Nat Med. 2001; 7(6):719-24. PMC: 7095854. DOI: 10.1038/89098. View

Lwamba H, Alvarez R, Wise M, Yu Q, Halvorson D, Njenga M . Comparison of the full-length genome sequence of avian metapneumovirus subtype C with other paramyxoviruses. Virus Res. 2004; 107(1):83-92. DOI: 10.1016/j.virusres.2004.07.002. View

RYAN K, Portner A, Murti K . Antibodies to paramyxovirus nucleoproteins define regions important for immunogenicity and nucleocapsid assembly. Virology. 1993; 193(1):376-84. DOI: 10.1006/viro.1993.1134. View

Barr J, Chambers P, Harriott P, Pringle C, Easton A . Sequence of the phosphoprotein gene of pneumonia virus of mice: expression of multiple proteins from two overlapping reading frames. J Virol. 1994; 68(8):5330-4. PMC: 236484. DOI: 10.1128/JVI.68.8.5330-5334.1994. View

10.

Garcia J, Vivo A, Melero J . Cytoplasmic inclusions of respiratory syncytial virus-infected cells: formation of inclusion bodies in transfected cells that coexpress the nucleoprotein, the phosphoprotein, and the 22K protein. Virology. 1993; 195(1):243-7. DOI: 10.1006/viro.1993.1366. View

11.

Ling R, Easton A, Pringle C . 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. 1992; 73 ( Pt 7):1709-15. DOI: 10.1099/0022-1317-73-7-1709. View

12.

Locke D, Sellers H, Crawford J, Schultz-Cherry S, King D, Meinersmann R . Newcastle disease virus phosphoprotein gene analysis and transcriptional editing in avian cells. Virus Res. 2000; 69(1):55-68. DOI: 10.1016/s0168-1702(00)00175-1. View

13.

Geysen H, Rodda S, Mason T, Tribbick G, Schoofs P . Strategies for epitope analysis using peptide synthesis. J Immunol Methods. 1987; 102(2):259-74. DOI: 10.1016/0022-1759(87)90085-8. View

14.

Seal B . Avian pneumoviruses and emergence of a new type in the United States of America. Anim Health Res Rev. 2001; 1(1):67-72. DOI: 10.1017/s1466252300000062. View

15.

Kolakofsky D, Pelet T, GARCIN D, Hausmann S, Curran J, Roux L . Paramyxovirus RNA synthesis and the requirement for hexamer genome length: the rule of six revisited. J Virol. 1998; 72(2):891-9. PMC: 124558. DOI: 10.1128/JVI.72.2.891-899.1998. View

16.

Curran J . A role for the Sendai virus P protein trimer in RNA synthesis. J Virol. 1998; 72(5):4274-80. PMC: 109657. DOI: 10.1128/JVI.72.5.4274-4280.1998. View

17.

Ahmadian G, Chambers P, Easton A . Detection and characterization of proteins encoded by the second ORF of the M2 gene of pneumoviruses. J Gen Virol. 1999; 80 ( Pt 8):2011-2016. DOI: 10.1099/0022-1317-80-8-2011. View

18.

Lewis J, Chang G, Lanciotti R, TRENT D . Direct sequencing of large flavivirus PCR products for analysis of genome variation and molecular epidemiological investigations. J Virol Methods. 1992; 38(1):11-23. DOI: 10.1016/0166-0934(92)90165-a. View

19.

Briedis D, Lamb R, Choppin P . Influenza B virus RNA segment 8 codes for two nonstructural proteins. Virology. 1981; 112(2):417-25. DOI: 10.1016/0042-6822(81)90289-0. View

20.

Peeters B, Verbruggen P, Nelissen F, de Leeuw O . The P gene of Newcastle disease virus does not encode an accessory X protein. J Gen Virol. 2004; 85(Pt 8):2375-2378. DOI: 10.1099/vir.0.80160-0. View