Richards O C, Hey T D, Ehrenfeld E
J Virol. 1987 Jul;61(7):2307-10. doi: 10.1128/JVI.61.7.2307-2310.1987.
A portion of poliovirus double-stranded RNA (25 to 50%) isolated from infected HeLa cells contains hairpin loops at one end of the duplex structure. These structures rapidly reformed double-stranded molecules after denaturation and appeared as molecules of up to two times genome length upon electrophoresis in denaturing agarose gels. A second form of poliovirus double-stranded RNA was readily denaturable into genome length strands. When the hairpin RNA was treated with S1 nuclease, subsequent denaturation resulted in formation of strands of up to genome length. Hairpin molecules contained very little, if any, poly(A) sequences, suggesting that the hairpin forms after nucleolytic removal of the 3' end of plus-strand templates. We conclude that the hairpin double-stranded RNA found in infected cells is likely generated by intracellular nicking and self-priming and that it does not represent an intermediate in the process of RNA replication.
从感染的HeLa细胞中分离出的一部分脊髓灰质炎病毒双链RNA(25%至50%)在双链结构的一端含有发夹环。这些结构在变性后迅速重新形成双链分子,并且在变性琼脂糖凝胶中电泳时呈现出长度达基因组两倍的分子。脊髓灰质炎病毒双链RNA的第二种形式很容易变性成长度为基因组的链。当用S1核酸酶处理发夹RNA时,随后的变性导致形成长度达基因组的链。发夹分子即使含有也只含有极少的聚(A)序列,这表明发夹是在对正链模板的3'端进行核酸酶解去除后形成的。我们得出结论,在感染细胞中发现的发夹双链RNA可能是由细胞内切口和自我引发产生的,并且它并不代表RNA复制过程中的中间体。