Activation defects caused by mutations in Escherichia coli rpoA are promoter specific.
Escherichia coli RNA polymerases containing mutated alpha subunits were tested for their ability to respond to three different positive regulators (activators) in vitro. The two alpha (rpoA) mutants, alpha-256 and alpha-235, have deletions of the C-terminal 73 and 94 amino acids, respectively. In runoff transcription assays catalyzed by reconstituted holoenzyme, the effects of the mutations on each of three promoters tested were different: activation of the lambda pRM promoter by cI protein (repressor) was nearly normal, activation of the lambda pRE promoter by cII protein was reduced approximately fivefold, and direct activation of the trpPB promoter of Pseudomonas aeruginosa was completely inhibited. We also found that the reconstituted mutant enzyme was defective in recognition of trpPI in the absence of activator. The differential responses of the three promoters to their activators in the presence of the mutant enzymes indicate that the location of an activator-binding site does not by itself determine the region of RNA polymerase with which the activator interacts.
Gussin
G N
GN
Biology Department, University of Iowa, Iowa City 52242.
Olson
C
C
Igarashi
K
K
Ishihama
A
A
D001665:Binding Sites; D012321:DNA-Directed RNA Polymerases / Q000235:genetics*; D004926:Escherichia coli / Q000235:genetics*; D015971:Gene Expression Regulation, Enzymologic; D009154:Mutation; D011401:Promoter Regions, Genetic; D039381:RNA Cap-Binding Proteins; D016601:RNA-Binding Proteins / Q000502:physiology
D039381:RNA Cap-Binding Proteins; D016601:RNA-Binding Proteins; D012321:DNA-Directed RNA Polymerases