RNA Replication: Required Intermediates and the Dissociation of Template, Product, and Qβ Replicase

Carl Dobkin, Donald R. Mills, Fred Russell Kramer, Sol Spiegelman

Research output: Contribution to journalArticle

30 Citations (Scopus)

Abstract

Replication complexes containing only one molecule of Qβ replicase and one strand of midivariant RNA (MDV-1 RNA) template were prepared by incubating the replicase with an excess of MDV-1 (-) RNA. In the presence of excess minus strands, these monoenzyme replication complexes were shown to synthesize essentially pure MDV-1 (+) RNA in both the first and second cycles of replication. When an equivalent concentration of mutant MDV-1 (-) RNA was added to this reaction before completion of the first cycle of replication, only wild-type MDV-1 (+) RNA was produced in the first cycle, but both mutant and wild-type MDV-1 (+) RNA were produced in the second cycle of replication. These results demonstrate that a monoenzyme complex is competent to synthesize RNA and, therefore, that a multienzyme replication complex is not a necessary intermediate of replication. The data also imply that after the completion of chain elongation, the product strand is released from the replication complex and that the template and the replicase then dissociate.

Original languageEnglish (US)
Pages (from-to)2038-2044
Number of pages7
JournalBiochemistry
Volume18
Issue number10
DOIs
StatePublished - Jan 1 1979

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RNA
Multienzyme Complexes
Elongation
Molecules

All Science Journal Classification (ASJC) codes

  • Biochemistry

Cite this

Dobkin, Carl ; Mills, Donald R. ; Kramer, Fred Russell ; Spiegelman, Sol. / RNA Replication : Required Intermediates and the Dissociation of Template, Product, and Qβ Replicase. In: Biochemistry. 1979 ; Vol. 18, No. 10. pp. 2038-2044.
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RNA Replication : Required Intermediates and the Dissociation of Template, Product, and Qβ Replicase. / Dobkin, Carl; Mills, Donald R.; Kramer, Fred Russell; Spiegelman, Sol.

In: Biochemistry, Vol. 18, No. 10, 01.01.1979, p. 2038-2044.

Research output: Contribution to journalArticle

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N2 - Replication complexes containing only one molecule of Qβ replicase and one strand of midivariant RNA (MDV-1 RNA) template were prepared by incubating the replicase with an excess of MDV-1 (-) RNA. In the presence of excess minus strands, these monoenzyme replication complexes were shown to synthesize essentially pure MDV-1 (+) RNA in both the first and second cycles of replication. When an equivalent concentration of mutant MDV-1 (-) RNA was added to this reaction before completion of the first cycle of replication, only wild-type MDV-1 (+) RNA was produced in the first cycle, but both mutant and wild-type MDV-1 (+) RNA were produced in the second cycle of replication. These results demonstrate that a monoenzyme complex is competent to synthesize RNA and, therefore, that a multienzyme replication complex is not a necessary intermediate of replication. The data also imply that after the completion of chain elongation, the product strand is released from the replication complex and that the template and the replicase then dissociate.

AB - Replication complexes containing only one molecule of Qβ replicase and one strand of midivariant RNA (MDV-1 RNA) template were prepared by incubating the replicase with an excess of MDV-1 (-) RNA. In the presence of excess minus strands, these monoenzyme replication complexes were shown to synthesize essentially pure MDV-1 (+) RNA in both the first and second cycles of replication. When an equivalent concentration of mutant MDV-1 (-) RNA was added to this reaction before completion of the first cycle of replication, only wild-type MDV-1 (+) RNA was produced in the first cycle, but both mutant and wild-type MDV-1 (+) RNA were produced in the second cycle of replication. These results demonstrate that a monoenzyme complex is competent to synthesize RNA and, therefore, that a multienzyme replication complex is not a necessary intermediate of replication. The data also imply that after the completion of chain elongation, the product strand is released from the replication complex and that the template and the replicase then dissociate.

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