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Nucleic Acids Research, 1977, Vol. 4, No. 4 827-842
© 1977

Articles

Transcription in vitro of bacteriophage lambda 4S RNA: studies on termination and rho protein

Bruce H. Howard, Benoit de Crombrugghe and Martin Rosenberg

Laboratory of Molecular Biology, National Cancer Institute National Institutes of Health Bethesda, MD 20014, USA

Received December 21, 1976. When bacteriophage {lambda}pgal8 DNA is transcribed in a purified in vitro system by E. coli RNA polymerase (nucleoside triphosphate: RNA nucleotidyl-transferase, EC 2.7.7.6 [EC] ), several major transcripts are synthesized. We have investigated transcriptional termination of one of these transcripts, the 4S, or "oop" RNA. Analysis by two-dimensional "fingerprinting" of Tl oligonucleotides reveals that transcription of the 4S RNA terminates at a specific site on the {lambda}pgal8 DNA template, tL' with an efficiency of approximately 80Z, i.e. 20% of transcripts are extended into larger RNAs. Addition of the E. coli protein rho to our transcription reactions has two effects: a) the efficiency of termination at the tL' site is increased to 100%; b) the number of AS transcripts synthesized is increased by greater than 5-fold. Rho appears to stimulate 4S RNA synthesis by facilitating more rapid release of RNA polymerase from the tL' termination site.


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M Rosenberg, A. Chepelinsky, and K McKenney
Studying promoters and terminators by gene fusion
Science, November 18, 1983; 222(4625): 734 - 739.
[Abstract] [PDF]


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