Nucleic Acids Research, 1995, Vol. 23, No. 5 827-834
© 1995
MOLECULAR BIOLOGY |
Promoter determinants for Escherichia coli RNA polymerase holoenzyme containing 
38 (the rpoS gene product)
Institute of Molecular and Cellular Biosciences, The University of Tokyo Bunkyo-ku, Tokyo 113, Japan Department of Molecular Genetics, National Institute of Genetics Mishima, Shizuoka 411, Japan
*To whom correspondence should be addressed
Received November 2, 1994. Revised January 20, 1995. Accepted January 20, 1995.
Sequence determinants responsible for promoter recognition by RNA polymerase holoenzyme containing 
38, the rpoS gene product, were analyzed. In a previous study [Tanaka et al. (1993) Proc. Natl. Acad. Sci. USA, 90, 3511-3515], Escherichia coli promoters were classified into three groups: promoters recognized only by RNA polymerase holoenzyme containing 70 (E70); promoters recognized preferentially by that containing 38 (E38); promoters recognized by both E70 and E38. As representatives of each group of promoter, we chose the alaS, fic and lacUV5 promoters. Making use of a restriction enzyme site inserted between the –10 and –35 hexamer sequences, promoters were divided into the upstream (UE) and downstream (DE) elements. These UEs and DEs were combined in all possible combinations and used for in vitro transcription reactions. Promoters containing DE from the fic or lacUV5 promoter were found to be recognized by E38, while those containing DE from the alaS promoter were not. Moreover, fic DE alone functioned as an efficient promoter for E38. Thus we conclude that the discrimination signal resides within the DE sequence. To test the activator response of E38, in vitro transcription reactions were also performed with the gal and lac promoters. For both CRP-responsive P1 promoters, E38 was found to be activated by the CRP-cAMP complex.
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