Cubitt B, Oldstone C, de la Torre J C
Department of Neuropharmacology, Scripps Research Institute, La Jolla, California 92037.
J Virol. 1994 Mar;68(3):1382-96. doi: 10.1128/JVI.68.3.1382-1396.1994.
We have previously demonstrated that Borna disease virus (BDV) has a negative nonsegmented single-stranded (NNS) RNA genome that replicates in the nucleus of infected cells. Here we report for the first time the cloning and complete sequence of the BDV genome. Our results revealed that BDV has a genomic organization similar to that of other members of the Mononegavirales order. We have identified five main open reading frames (ORFs). The largest ORF, V, is located closest to the 5' end in the BDV genome and, on the basis of strong homology with other NNS-RNA virus polymerases, is a member of the L-protein family. The intercistronic regions vary in length and nucleotide composition and contain putative transcriptional start and stop signals. BDV untranslated 3' and 5' RNA sequences resemble those of other NNS-RNA viruses. Using a set of overlapping probes across the BDV genome, we identified nine in vivo synthesized species of polyadenylated subgenomic RNAs complementary to the negative-strand RNA genome, including monocistronic transcripts corresponding to ORFs I, II, and IV, as well as six polycistronic polyadenylated BDV RNAs. Interestingly, although ORFs III and V were detected within polycistronic transcripts, their corresponding monocistronic transcripts were not detected. Our data indicate that BDV is a member of the Mononegavirales, specially related to the family Rhabdoviridae. However, in contrast to the rest of the NNS-RNA animal viruses, BDV replication and transcription occur in the nucleus of infected cells. These findings suggest a possible relationship between BDV and the plant rhabdoviruses, which also replicate and transcribe in the nucleus.
我们之前已经证明，博尔纳病病毒（BDV）具有负链非节段单链（NNS）RNA基因组，该基因组在受感染细胞的细胞核中复制。在此，我们首次报告了BDV基因组的克隆及完整序列。我们的结果显示，BDV的基因组结构与单股负链RNA病毒目其他成员相似。我们鉴定出了五个主要的开放阅读框（ORF）。最大的ORF，即ORF V，位于BDV基因组中最靠近5'端的位置，基于与其他NNS-RNA病毒聚合酶的高度同源性，它是L蛋白家族的成员。基因间隔区的长度和核苷酸组成各不相同，并包含假定的转录起始和终止信号。BDV的3'和5'非翻译RNA序列与其他NNS-RNA病毒的相似。使用一组跨越BDV基因组的重叠探针，我们鉴定出了九种在体内合成的与负链RNA基因组互补的多聚腺苷酸化亚基因组RNA，包括与ORF I、II和IV对应的单顺反子转录本，以及六种多顺反子多聚腺苷酸化BDV RNA。有趣的是，尽管在多顺反子转录本中检测到了ORF III和V，但未检测到它们相应的单顺反子转录本。我们的数据表明，BDV是单股负链RNA病毒目的成员，特别与弹状病毒科相关。然而，与其他NNS-RNA动物病毒不同的是，BDV的复制和转录发生在受感染细胞的细胞核中。这些发现提示了BDV与植物弹状病毒之间可能存在的关系, 植物弹状病毒也在细胞核中进行复制和转录。
