期刊
EMBO JOURNAL
卷 19, 期 1, 页码 124-133出版社
WILEY
DOI: 10.1093/emboj/19.1.124
关键词
bacteriophage phi 6; double-stranded RNA virus; purified RNA-dependent RNA polymerase; replication in vitro
In nature, synthesis of both minus- and plus-sense RNA strands of all the known double-stranded RNA viruses occurs in the interior of a large protein assembly referred to as the polymerase complex. In addition to other proteins, the complex contains a putative polymerase possessing characteristic sequence motifs, However, none of the previous studies has shown template-dependent RNA synthesis directly with an isolated putative polymerase protein. In this report, recombinant protein P2 of double-stranded RNA bacteriophage phi 6 was purified and demonstrated in an in vitro enzymatic assay to act as the replicase, The enzyme efficiently utilizes phage-specific, positive-sense RNA substrates to produce double-stranded RNA molecules, which are formed by newly synthesized, full-length minus-strands base paired with the plus: strand templates. P2-catalyzed replication is also shown to be very effective with a broad range of heterologous single-stranded RNA templates. The importance and implications of these results are discussed.
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