Identification and characterization of three immunodominant structural proteins of fowlpox virus
Identification and characterization of three immunodominant structural proteins of fowlpox virus
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DOI：
10.1128/jvi.76.19.9844-9855.2002
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发表时间：
2002-10-01
影响因子：
5.4
通讯作者：
Skinner, MA
中科院分区：
文献类型：
--
作者：
Boulanger, D;Green, P;Skinner, MA
鸡痘病毒（FWPV）结构蛋白基因的鉴定主要是通过与牛痘病毒（VACV）结构蛋白基因的序列同源性来进行的，但对FWPV结构蛋白的编码基因的研究却很少。针对Poxine和HP 1 -440（慕尼黑）克隆FP 9的单克隆抗体（MAbs）的生产允许鉴定三种免疫显性FWPV蛋白：39-kDa核心蛋白（由FPV 168编码，与VACV A4 L同源），30-和35-kDa蛋白质双联体，以及丰富的63-kDa蛋白质。30-和35-kDa蛋白是非糖基化的抗原相关蛋白，存在于细胞内成熟病毒膜中，并与病毒工厂紧密定位。N-末端测序鉴定出35-kDa蛋白由FPV 140（VACV H3 L的FWPV同系物）编码。63-kDa蛋白质形成共价连接的二聚体和寡聚体。它仍然主要是不溶的洗涤剂处理纯化的病毒，但没有本地化密切与病毒工厂。N-末端测序不成功，表明N-末端封闭。CNBr消化产生了由FPV 191编码的肽，预测其编码两种FWPV A型包涵体（ATI）蛋白中的一种。63-kDa蛋白的特征与发表的关于牛痘或VACV ATI蛋白的观察结果不一致（它似乎是必需的）。然而，63-kDa蛋白与VACV p4 c病毒封闭和14-kDa融合蛋白具有共同特征。痘病毒ATI基因座（VACV A24 R和A28 L之间）的基因分配因序列冗余和变异而复杂化，通常是由于缺失和多个移码突变。FPV 191在这个位点的基因的身份进行了讨论。
Genes encoding fowlpox virus (FWPV) structural proteins have been identified mainly by sequence homology with those from vaccinia virus (VACV), but little is known about the encoded proteins. Production of monoclonal antibodies (MAbs) against Poxine and HP1-440 (Munich) clone FP9 allowed the identification of three immunodominant FWPV proteins: the 39-kDa core protein (encoded by FPV168,homologous to VACV A4L), a 30- and 35-kDa protein doublet, and an abundant 63-kDa protein. The 30- and 35-kDa proteins are nonglycosylated, antigenically related proteins present in the intracellular mature virus membrane and localizing closely with the viral factories. N-terminal sequencing identified the 35-kDa protein as encoded by FPV140 (the FWPV homolog of VACV H3L). The 63-kDa protein forms covalently linked dimers and oligomers. It remained mainly insoluble upon detergent treatment of purified virus but did not localize closely with the viral factory. N-terminal sequencing was unsuccessful, suggesting N-terminal blocking. CNBr digestion generated a peptide encoded by FPV191, predicted to encode one of two FWPV A-type inclusion (ATI) proteins. The characteristics of the 63-kDa protein were inconsistent with published observations on cowpox or VACV ATI proteins (it appears to be essential). The 63-kDa protein, however, shares characteristics with both VACV p4c virus occlusion and 14-kDa fusion proteins. Gene assignment at the poxvirus ATI locus (between VACV A24R and A28L) is complicated by sequence redundancies and variations, often due to deletions and multiple frameshift mutations. The identity of FPV191 in relation to genes at this locus is discussed.