Ewen M E, Revel H R
Department of Molecular Genetics and Cell Biology, University of Chicago, Illinois 60637.
Virology. 1990 Oct;178(2):509-19. doi: 10.1016/0042-6822(90)90348-u.
RNA-protein complexes active for transcription and replication of the double-stranded RNA bacteriophage phi 6 have been partially purified from lysates of infected Pseudomonas phaseolicola. Transcribing particles (filled procapsids) contain the three viral dsRNAs and all four procapsid proteins P1, P2, P4, and P7. Particles with replicase activity contain the same four proteins as well as single plus RNA strands duplexed with various extents of minus strands initiated in vivo. The in vitro replication reaction is insensitive to RNaseA. Sarkosyl destroys transcription complexes but does not reduce the activity of replication complexes, although the latter lose 80% of their P4 and the single-strand RNA template becomes sensitive to RNase. The detection of complexes that replicate small only, or both small and medium, RNA suggests that the RNAs are packaged sequentially in the order small, medium, large.
对双链RNA噬菌体φ6转录和复制具有活性的RNA-蛋白质复合物已从感染菜豆假单胞菌的裂解物中部分纯化出来。转录颗粒(完整的原衣壳)包含三种病毒双链RNA以及所有四种原衣壳蛋白P1、P2、P4和P7。具有复制酶活性的颗粒含有相同的四种蛋白质以及与体内起始的不同程度的负链双链化的单正链RNA。体外复制反应对核糖核酸酶A不敏感。十二烷基肌氨酸钠会破坏转录复合物,但不会降低复制复合物的活性,尽管后者会失去80%的P4,并且单链RNA模板对核糖核酸酶变得敏感。对仅复制小RNA或同时复制小RNA和中RNA的复合物的检测表明,RNA按小、中、大的顺序依次包装。