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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
. 1978 Jul;75(7):3220–3224. doi: 10.1073/pnas.75.7.3220

Novel template requirements of N4 virion RNA polymerase.

S C Falco, R Zivin, L B Rothman-Denes
PMCID: PMC392746  PMID: 356050

Abstract

A DNA-dependent RNA polymerase has been purified from disrupted virions of the Escherichia coli bacteriophage N4. The RNA polymerase is phage-coded and is required for class I N4 RNA synthesis, which is defined as RNA synthesized in vivo in the absence of post-infection protein synthesis. A polypeptide of molecular weight 350,000 is detected when the purified enzyme is analyzed by sodium dodecyl sulfate/polyacrylamide gel electrophoresis. N4 RNA polymerase requires denatured DNA as a template in vitro and shows a strong preference for denatured N4 DNA. With this template, transcription is asymmetric. The RNA product is complementary to only the H strand of N4 DNA. Furthermore, only class I N4 RNA is synthesized. In vivo transcription by the N4 virion RNA polymerase is inhibited by coumermycin. This result suggests that the activity of E. coli DNA gyrase, an enzyme that introduces negative supertwists into DNA, is required for N4 transcription.

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Selected References

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