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Journal of Clinical Microbiology, April 2003, p. 1730-1735, Vol. 41, No. 4
0095-1137/03/$08.00+0 DOI: 10.1128/JCM.41.4.1730-1735.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.
Nucleotide and Predicted Amino Acid Sequence-Based Analysis of the Avian Metapneumovirus Type C Cell Attachment Glycoprotein Gene: Phylogenetic Analysis and Molecular Epidemiology of U.S. Pneumoviruses
Rene Alvarez,1 Humphrey M. Lwamba,2 Darrell R. Kapczynski,1 M. Kariuki Njenga,2 and Bruce S. Seal1*Southeast Poultry Research Laboratory, Agricultural Research Service, U.S. Department of Agriculture, Athens, Georgia 30605,1 Department of Veterinary Pathobiology, University of Minnesota, St. Paul, Minnesota 551082
Received 3 September 2002/ Returned for modification 16 December 2002/ Accepted 23 January 2003
A serologically distinct avian metapneumovirus (aMPV) was isolated in the United States after an outbreak of turkey rhinotracheitis (TRT) in February 1997. The newly recognized U.S. virus was subsequently demonstrated to be genetically distinct from European subtypes and was designated aMPV serotype C (aMPV/C). We have determined the nucleotide sequence of the gene encoding the cell attachment glycoprotein (G) of aMPV/C (Colorado strain and three Minnesota isolates) and predicted amino acid sequence by sequencing cloned cDNAs synthesized from intracellular RNA of aMPV/C-infected cells. The nucleotide sequence comprised 1,321 nucleotides with only one predicted open reading frame encoding a protein of 435 amino acids, with a predicted Mr of 48,840. The structural characteristics of the predicted G protein of aMPV/C were similar to those of the human respiratory syncytial virus (hRSV) attachment G protein, including two mucin-like regions (heparin-binding domains) flanking both sides of a CX3C chemokine motif present in a conserved hydrophobic pocket. Comparison of the deduced G-protein amino acid sequence of aMPV/C with those of aMPV serotypes A, B, and D, as well as hRSV revealed overall predicted amino acid sequence identities ranging from 4 to 16.5%, suggesting a distant relationship. However, G-protein sequence identities ranged from 72 to 97% when aMPV/C was compared to other members within the aMPV/C subtype or 21% for the recently identified human MPV (hMPV) G protein. Ratios of nonsynonymous to synonymous nucleotide changes were greater than one in the G gene when comparing the more recent Minnesota isolates to the original Colorado isolate. Epidemiologically, this indicates positive selection among U.S. isolates since the first outbreak of TRT in the United States.
* Corresponding author. Mailing address: Southeast Poultry Research Laboratory, ARS, USDA, 934 College Station Rd., Athens, GA 30605. Phone: (706) 546-3463. Fax: (706) 546-3161. E-mail: bseal{at}seprl.usda.gov.
Journal of Clinical Microbiology, April 2003, p. 1730-1735, Vol. 41, No. 4
0095-1137/03/$08.00+0 DOI: 10.1128/JCM.41.4.1730-1735.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.
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