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Nucleic Acids Research, 1984, Vol. 12, No. 2 1287-1299
© 1984

MOLECULAR BIOLOGY

A potential stem-loop structure and the sequence CAAUCAA in the transcript are insufficient to signal {varrho}-dependent transcription termination at {lambda}tR1

Lester F. Lau*,§, Jeffrey W. Roberts*, Ray WU*, Fawzy Georges+ and Saran A. Narang+

*Section of Biochemistry, Molecular and Cell Biology, Cornell University Ithaca, NY 14853, USA +Division of Biological Sciences, National Research Council of Canada Ottawa KIA OR6, Canada

Received August 11, 1983. Revised November 2, 1983. Accepted November 16, 1983.

It has been suggested that a sequence in the RNA transcript that can form a stem and loop structure, followed by the sequence CAAUCAA, is the signal for {rho}-dependent transcription termination. We tested this hypothesis by synthesizing a DNA duplex whose sequence corresponds to a region of the {lambda}tR1 terminator that contains these structural features. We cloned this synthehc DNA fragment under the control of the lacUV5 promoter, and showed that it does not cause {rho}-dependent termination in vitro RNA polymerase pauses during in vitro transcription across the synthetic sequence, although less efficiently than at the corresponding sequence on the {lambda} template. No {rho}-mediated termination was detected even under conditions that prolonged transcriptional pausing at the synthetic site, indicating that the synthetic sequence is defective as a transcript release site. We suggest that unlike {rho}-independent terminators, {rho}-dependent terminators require sequences in addition to those immediately before the sites of termination.

§Present address: Department of Molecular Biology and Genetics, Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA


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