Nucleic Acids Research, Vol 25, Issue 2 326-332, Copyright © 1997 by Oxford University Press
VA Rhodius, DM West, CL Webster, SJW Busby and NJ Savery
Transcription activation by the Escherichia coli cyclic AMP receptor
protein (CRP) at Class II promoters is dependent on direct interactions
between two surface-exposed activating regions (AR1 and AR2) and two
contact sites in RNA polymerase. The effects on transcription activation of
disrupting either AR1 or AR2 have been measured at different Class II
promoters. AR2 but not AR1 is essential for activation at all the Class II
promoters that were tested. The effects of single positive control
substitutions in AR1 and AR2 vary from one promoter to another: the effects
of the different substitutions are contingent on the -35 hexamer sequence.
Abortive initiation assays have been used to quantify the effects of
positive control substitutions in each activating region on the kinetics of
transcription initiation at the Class II CRP- dependent promoter pmelRcon.
At this promoter, the HL159 substitution in AR1 results in a defect in the
initial binding of RNA polymerase whilst the KE101 substitution in AR2
reduces the rate of isomerization from the closed to the open complex.
ARTICLES
Transcription activation at class II CRP-dependent promoters: the role of different activating regions
School of Biochemistry, University of Birmingham, PO Box 363, Birmingham B15 2TT, UK.
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