The genome length of human parainfluenza virus type 2 follows the rule of six, and recombinant viruses recovered from non-polyhexameric-length antigenomic cDNAs contain a biased distribution of correcting mutations
The genome length of human parainfluenza virus type 2 follows the rule of six, and recombinant viruses recovered from non-polyhexameric-length antigenomic cDNAs contain a biased distribution of correcting mutations
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DOI：
10.1128/jvi.77.1.270-279.2003
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发表时间：
2003-01-01
影响因子：
5.4
通讯作者：
Murphy, BR
中科院分区：
文献类型：
--
作者：
Skiadopoulos, MH;Vogel, L;Murphy, BR
副粘病毒科病毒的副粘病毒亚科成员有一个不寻常的要求，即病毒基因组的核苷酸长度必须是六的偶数倍，以便进行有效的RNA复制，从而发生病毒复制。人类副流感病毒2型（HPIV2）是该属中唯一被报道基因组长度不是六的偶数倍的成员，而且它也是从不符合“六规则”的全长反基因组正义cDNA中回收的。为了重新检查 HPIV2 天然分离株的核苷酸长度问题，确定了三种 HPIV2 毒株的完整共有基因组序列：Greer、Vanderbilt/1994 (V94) 和 Vanderbilt/1998。这些菌株中的每一个被发现具有 15,654 个核苷酸 (nt) 的基因组长度，因此在每种情况下都符合六的规则。为了直接检验 HPIV2 基因组长度为 6 的偶数倍的要求，我们构建了 6 个全长反基因组 HPIV2/V94 cDNA，其与多六聚体长度偏差 0 至 5 nt。重组 HPIV2 很容易从所有 cDNA 中回收，包括那些不符合 6 规则的 cDNA。对一种重组 HPIV2 分离株的每种非多聚六聚体反基因组 cDNA 进行了完整测序。发现它们含有小的核苷酸插入或缺失，赋予恢复的基因组多六聚体长度。有趣的是，几乎所有的长度校正都发生在血凝素-神经氨酸酶和大聚合酶基因或插入的基因间区域内，因此最接近导致偏离六规则的插入片段。这些结果表明，在完全感染性病毒的背景下，HPIV2 对多六聚体基因组具有强烈且看似绝对的需求。
Members of the Paramyxovirinae subfamily of the Paramyxoviridae family of viruses have the unusual requirement that the nucleotide length of the viral genome must be an even multiple of six in order for efficient RNA replication, and hence virus replication, to occur. Human parainfluenza virus type 2 (HPIV2) is the only member of the genus that has been reported to have a genome length that is not an even multiple of six, and it has also been recovered from a full-length antigenomic-sense cDNA that did not conform to the "rule of six." To reexamine the issue of nucleotide length in natural isolates of HPIV2, a complete consensus genomic sequence was determined for three HPIV2 strains: Greer, Vanderbilt/1994 (V94), and Vanderbilt/1998. Each of these strains was found to have a genome length of 15,654 nucleotides (nt), thus conforming in each case to the rule of six. To directly examine the requirement that the genomic length of HPIV2 be an even multiple of six, we constructed six full-length antigenomic HPIV2/V94 cDNAs that deviated from a polyhexameric length by 0 to 5 nt. Recombinant HPIV2s were readily recovered from all of the cDNAs, including those that did not conform to the rule of six. One recombinant HPIV2 isolate was completely sequenced for each of the nonpolyhexameric antigenomic cDNAs. These were found to contain small nucleotide insertions or deletions that conferred polyhexameric length to the recovered genome. Interestingly, almost all of the length corrections occurred within the hemagglutinin-neuraminidase and large polymerase genes or the intervening intergenic region and thus were proximal to the insert that caused the deviation from the rule of six. These results demonstrate, in the context of complete infectious virus, that HPIV2 has a strong and seemingly absolute requirement for a polyhexameric genome.