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Nucleic Acids Research, 1989, Vol. 17, No. 10 3927-3949
© 1989

MOLECULAR BIOLOGY

Pseudorevertants of a lac promoter mutation reveal overlapping nascent promoters

Russell Karls, Vincent Schulz, Stevan B. Jovanovich1, Sheila Flynn+,, Alex Pak§, and William S. Reznikoff*,

1McArdle Laboratory for Cancer Research, University of Wisconsin—Madison Madison, WI 53706, USA Department of Biochemistry, College of Agricultural and Life Sciences Madison, WI 53706, USA

*To whom correspondence should be addressed

Received December 16, 1988. Revised April 21, 1989. Accepted April 21, 1989.

Four pseudorevertants of a –10 region lacPmutation were isolated. Three of these mutations were found to activate nascent promoters. These mutations were: a –2 G/C -> A/T change (–2A) promoting transcription at position +11, a +1 A/T -> T/A change (+1T) promoting transcription initiation at position +13, and a +10 C/G -> A/T change (+1OA) promoting transcription initiation at a complex series of positions. The fourth mutation [a –12 T/A -> A/T change (–12A)] promotes transcription initiation at –1. The promoters activated by mutations –12A, –2A and +1T resembled the canonical sigma 70 promoter sequences. The +1OA promoter activity is also dependent upon the sigma 70 holoenzyme but can not be readily assigned to a specific promoter sequence.

+Present addresses: University of Minnesota Medical School, Rochester, MN

§Present addresses: University of Wisconsin Medical School, Madison, WI, USA


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