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J Biol Chem, Vol. 273, Issue 30, 18802-18811, July 24, 1998

Characterization of an Unusual, Sequence-specific Termination Signal for T7 RNA Polymerase

Biao HeDagger , Alexander KukarinDagger parallel , Dmitry TemiakovDagger , Stephen T. Chin-BowDagger , Dmitry L. LyakhovDagger **, Minqing RongDagger , Russell K. DurbinDagger , and William T. McAllisterDagger

From the Dagger  Department of Microbiology and Immunology, Morse Institute for Molecular Genetics, State University of New York, Health Science Center, Brooklyn, New York 11203-2098, the parallel  Laboratory for Molecular Genetics of Microorganisms, Institute of Molecular Genetics, Russian Academy of Sciences, 46 Kurchatov Square, Moscow 123182, Russia, and the ** V.A. Engelhardt Institute of Molecular Biology, Russian Academy of Sciences, 32 Vavilov St., Moscow 117984, Russia

We have characterized an unusual type of termination signal for T7 RNA polymerase that requires a conserved 7-base pair sequence in the DNA (ATCTGTT in the non-template strand). Each of the nucleotides within this sequence is critical for function, as any substitutions abolish termination. The primary site of termination occurs 7 nucleotides downstream from this sequence but is context-independent (that is, the sequence around the site of termination, and in particular the nucleotide at the site of termination, need not be conserved). Termination requires the presence of the conserved sequence and its complement in duplex DNA and is abolished or diminished if the signal is placed downstream of regions in which the non-template strand is missing or mismatched. Under the latter conditions, much of the RNA product remains associated with the template. The latter results suggest that proper resolution of the transcription bubble at its trailing edge and/or displacement of the RNA product are required for termination at this class of signal.

Copyright © 1998 by The American Society for Biochemistry and Molecular Biology, Inc.
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