Nucleic Acids Research, 1981, Vol. 9, No. 18 4689-4707
© 1981
MOLECULAR BIOLOGY |
Termination of transcription of the coliphage T7 "early" operon in vitro: slowness of enzyme release, and lack of any role for sigma
Department of Molecular Biology, University of Edinburgh Scotland, UK
Received June 30, 1981. The leftmost portion of the coliphage T7 genome is transcribed by the RNA polymerase Escherichia coli immediately after infection. This "early" operon is delineated by three promoters on the left, and a transcriptional terminator on the right. The terminator is efficient both in vivo, and with highly purified RNA polymerase in vitro. We have studied termination in vitro, using synchronously initiated transcription reactions with low enzyme:DNA ratios. We show that recognition of the stop signal and release of RNA product are relatively rapid. Dissociation of the enzyme from the DNA is quite slow, and probably rate-limiting for re-cycling of the polymerase.
It is well established that the sigma subunit of RNA polymerase is required for specific initiation, but redundant during RNA elongation. By exploiting antisigma antiserum we have obtained evidence that sigma is also redundant during all steps of termination in vitro.