基于核苷酸和预测氨基酸序列的C型禽偏肺病毒细胞附着糖蛋白基因分析:美国肺病毒的系统发育分析和分子流行病学

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.

作者信息

Alvarez Rene, Lwamba Humphrey M, Kapczynski Darrell R, Njenga M Kariuki, Seal Bruce S

机构信息

Southeast Poultry Research Laboratory, Agricultural Research Service, U.S. Department of Agriculture, Athens, Georgia 30605, USA.

出版信息

J Clin Microbiol. 2003 Apr;41(4):1730-5. doi: 10.1128/JCM.41.4.1730-1735.2003.

Abstract

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 M(r) 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.

摘要

1997年2月美国爆发火鸡鼻气管炎(TRT)后,分离出一种血清学上不同的禽偏肺病毒(aMPV)。随后证明这种新发现的美国病毒在基因上与欧洲亚型不同,被指定为aMPV血清型C(aMPV/C)。我们测定了aMPV/C(科罗拉多毒株和三株明尼苏达分离株)编码细胞附着糖蛋白(G)的基因的核苷酸序列,并通过对从aMPV/C感染细胞的细胞内RNA合成的克隆cDNA进行测序预测了氨基酸序列。核苷酸序列由1321个核苷酸组成,只有一个预测的开放阅读框,编码一个435个氨基酸的蛋白质,预测分子量为48840。预测的aMPV/C的G蛋白的结构特征与人类呼吸道合胞病毒(hRSV)附着G蛋白的结构特征相似,包括在保守疏水口袋中存在的CX3C趋化因子基序两侧的两个粘蛋白样区域(肝素结合域)。将aMPV/C推导的G蛋白氨基酸序列与aMPV血清型A、B和D以及hRSV的序列进行比较,发现总体预测氨基酸序列同一性在4%至16.5%之间,表明它们之间关系较远。然而,当将aMPV/C与aMPV/C亚型内的其他成员进行比较时,G蛋白序列同一性在72%至97%之间,与最近鉴定的人类MPV(hMPV)G蛋白的同一性为21%。将最近的明尼苏达分离株与原始的科罗拉多分离株进行比较时,G基因中非同义核苷酸变化与同义核苷酸变化的比率大于1。从流行病学角度来看,这表明自美国首次爆发TRT以来,美国分离株之间存在正选择。

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