A genetic method for dissecting the mechanism of transcriptional activator synergy by identical activators.
Proc Natl Acad Sci U S A
; 96(22): 12673-8, 1999 Oct 26.
Article
em En
| MEDLINE
| ID: mdl-10535981
ABSTRACT
Pairs of transcriptional activators in prokaryotes have been shown to activate transcription synergistically from promoters with two activator binding sites. In some cases, such synergistic effects result from cooperative binding, but in other cases each DNA-bound activator plays a direct role in the activation process by interacting simultaneously with separate surfaces of RNA polymerase. In such cases, each DNA-bound activator must possess a functional activating region, the surface that mediates the interaction with RNA polymerase. When transcriptional activation depends on two or more identical activators, it is not straightforward to test the requirement of each activator for a functional activating region. Here we describe a method for directing a mutationally altered activator to either one or the other binding site, and we demonstrate the use of this method to examine the mechanism of transcriptional activator synergy by the Escherichia coli cyclic AMP receptor protein (CRP) working at an artificial promoter bearing two CRP-binding sites.
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Base de dados:
MEDLINE
Assunto principal:
Transativadores
Idioma:
En
Ano de publicação:
1999
Tipo de documento:
Article