Esteban R, Fujimura T, Wickner R B
Section on Genetics of Simple Eukaryotes, National Institute of Diabetes and Digestive and Kidney Diseases, Bethesda, MD 20892.
Proc Natl Acad Sci U S A. 1988 Jun;85(12):4411-5. doi: 10.1073/pnas.85.12.4411.
X double-stranded RNA is a deletion mutant of L-A double-stranded RNA and is encapsidated in viral particles by the L-A-encoded major coat protein. X double-stranded RNA has all the cis sites necessary to be transcribed, encapsidated, and replicated. We have cloned X double-stranded RNA and sequenced it. The complete X double-stranded RNA sequence deduced indicates that the first 25 bases of the X plus-strand 5' end originated from the 5' end of the L-A plus strand and that most, if not all, of the rest comes from the 3' end of the L-A plus strand. The X plus strand made by X double-stranded RNA-containing virus-like particles binds specifically to empty open virus-like particles and is converted by these particles to X double-stranded RNA. RNA transcripts of the X complementary DNA clones and deletion derivatives thereof were made in vitro by T7 and T3 RNA polymerases and tested for specific binding to the virus-like particles. The results suggest that the binding is due to the sequence UUUGGCCAGG, 370 bases upstream from the X plus-strand 3' end. This sequence is also present in the M1 plus strand 140 bases from its 3' end.
X双链RNA是L-A双链RNA的缺失突变体,由L-A编码的主要衣壳蛋白包裹在病毒颗粒中。X双链RNA具有转录、包裹和复制所需的所有顺式作用位点。我们已克隆X双链RNA并对其进行测序。推导得到的完整X双链RNA序列表明,X正链5'端的前25个碱基源自L-A正链的5'端,其余部分(如果不是全部)大多来自L-A正链的3'端。含X双链RNA的病毒样颗粒产生的X正链与空的开放病毒样颗粒特异性结合,并被这些颗粒转化为X双链RNA。通过T7和T3 RNA聚合酶在体外制备X互补DNA克隆及其缺失衍生物的RNA转录本,并测试其与病毒样颗粒的特异性结合。结果表明,这种结合是由于X正链3'端上游370个碱基处的序列UUUGGCCAGG。该序列也存在于M1正链3'端140个碱基处。