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PMID:10619851
| Citation |
Makeyev, EV and Bamford, DH (2000) Replicase activity of purified recombinant protein P2 of double-stranded RNA bacteriophage phi6. EMBO J. 19:124-33 |
|---|---|
| Abstract |
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 phi6 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. |
| Links |
PubMed PMC1171784 Online version:10.1093/emboj/19.1.124 |
| Keywords |
Amino Acid Sequence; Bacteriophage phi 6/enzymology; DNA-Directed DNA Polymerase/metabolism; Electrophoresis, Agar Gel; Escherichia coli; Molecular Sequence Data; RNA Replicase/isolation & purification; RNA Replicase/metabolism; RNA, Double-Stranded/metabolism; Recombinant Proteins/metabolism; Ribonucleases/metabolism; Templates, Genetic |
| edit table |
Significance
Annotations
| Gene product | Qualifier | GO Term | Evidence Code | with/from | Aspect | Extension | Notes | Status |
|---|---|---|---|---|---|---|---|---|
| GO:0003968: RNA-directed RNA polymerase activity |
ECO:0000314: |
F |
Figure 2 "Recombinant P2 catalyzes RNA-dependent RNA synthesis in vitro." |
complete | ||||
|
enables |
GO:0003968: RNA-directed 5'-3' RNA polymerase activity |
ECO:0000314: direct assay evidence used in manual assertion |
F |
Seeded From UniProt |
complete | |||
See also
References
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