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Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1972 Sep;69(9):2691–2695. doi: 10.1073/pnas.69.9.2691

Initiation of DNA Synthesis: Synthesis of ϕX174 Replicative Form Requires RNA Synthesis Resistant to Rifampicin*

Randy Schekman 1, William Wickner 1, Ole Westergaard 1, Douglas Brutlag 1, Klaus Geider 1, Leroy L Bertsch 1, Arthur Kornberg 1
PMCID: PMC427018  PMID: 4560696

Abstract

Conversion of single-stranded DNA of phage ϕX174 to the double-stranded replicative form in Escherichia coli uses enzymes essential for initiation and replication of the host chromosome. These enzymes can now be purified by the assay that this phage system provides. The ϕX174 conversion is distinct from that of M13. The reaction requires different host enzymes and is resistant to rifampicin and streptolydigin, inhibitors of RNA polymerase. However, RNA synthesis is essential for ϕX174 DNA synthesis: the reaction is inhibited by low concentrations of actinomycin D, all four ribonucleoside triphosphates are required, and an average of one phosphodiester bond links DNA to RNA in the isolated double-stranded circles. Thus, we presume that, as in the case of M13, synthesis of a short RNA chain primes the synthesis of a replicative form by DNA polymerase. Initiation of DNA synthesis by RNA priming is a mechanism of wide significance.

Keywords: dnaA gene product, dnaB gene product, M13 DNA, spermidine

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

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