Quackenbush S L, Holzschu D L, Bowser P R, Casey J W
Department of Microbiology and Immunology, Cornell University, Ithaca, New York 14853, USA.
Virology. 1997 Oct 13;237(1):107-12. doi: 10.1006/viro.1997.8755.
Walleye dermal sarcoma virus (WDSV) is a complex retrovirus associated with dermal sarcomas of walleye that develop and regress on a seasonal basis. WDSV contains, in addition to gag, pol, and env, three open reading frames (ORFs) designated ORF A, ORF B, and ORF C. The polymerase chain reaction technique was used to amplify and clone cDNAs representing subgenomic viral mRNAs isolated from developing (fall) and regressing (spring) tumors. Nine different singly or multiply spliced viral transcripts were identified and all were found to utilize a common 5' leader sequence. This leader sequence is spliced to the pol/env junction or downstream of env to generate singly spliced transcripts. Multiply spliced transcripts contain the 5' leader, the pol/env junction, and sequences derived from the 3' end of the genome. One multiply spliced transcript was isolated with the potential to encode the full-length ORF A protein. In addition, WDSV produced mRNAs that utilize alternative splice acceptor sites which would allow synthesis of five variant forms of the ORF A protein. In contrast, the ORF B protein is postulated to arise from a singly spliced transcript with the potential to encode the entire open reading frame. Spliced subgenomic transcripts representing ORF C mRNAs were not identified, suggesting that ORF C may be encoded from the full-length viral genomic transcript. We estimate that at least a 100-fold lower amount of the accessory/regulatory subgenomic transcripts exists in developing vs regressing tumors. These results demonstrate that WDSV undergoes an elaborate pattern of mRNA splicing similar to that of other complex retroviruses.
大眼狮鲈皮肤肉瘤病毒(WDSV)是一种复杂的逆转录病毒,与大眼狮鲈的皮肤肉瘤有关,这些肉瘤会随季节发展和消退。除gag、pol和env外,WDSV还包含三个开放阅读框(ORF),分别命名为ORF A、ORF B和ORF C。采用聚合酶链反应技术扩增并克隆代表从发育中的(秋季)和消退中的(春季)肿瘤中分离出的亚基因组病毒mRNA的cDNA。鉴定出九种不同的单剪接或多剪接病毒转录本,发现它们都利用一个共同的5'前导序列。该前导序列与pol/env连接点或env下游进行剪接,以产生单剪接转录本。多剪接转录本包含5'前导序列、pol/env连接点以及源自基因组3'端的序列。分离出一种有潜力编码全长ORF A蛋白 的多剪接转录本。此外,WDSV产生的mRNA利用了可变剪接受体位点,这将允许合成ORF A蛋白的五种变体形式。相比之下,推测ORF B蛋白来自一种有潜力编码整个开放阅读框的单剪接转录本。未鉴定出代表ORF C mRNA的剪接亚基因组转录本,这表明ORF C可能由全长病毒基因组转录本编码。我们估计,与消退中的肿瘤相比,发育中的肿瘤中辅助/调节亚基因组转录本的数量至少低100倍。这些结果表明,WDSV经历了一种与其他复杂逆转录病毒相似的精细mRNA剪接模式。