Activation of ara operons by a truncated AraC protein does not require inducer.
The araC gene of Escherichia coli encodes a protein that binds the inducer L-arabinose to activate the transcription of three ara operons. In a study to determine the functional domains within the AraC protein, we have generated a set of overlapping deletions from the proximal end of the araC gene. We found that the removal of up to nearly 60% of the coding sequence of this protein still allows transcriptional activation of the ara operons in vivo, up to 27% that of the wild type. These truncated proteins, however, no longer require arabinose for induction. The ligand-induced conformational change apparently either releases or unmasks an existing functional domain within AraC, rather than generating a new conformation that is required for activation of the promoter of araBAD. Since the truncated protein of the mutant C154 (which lacks 153 amino acid residues from the N terminus) retains DNA binding specificity, the DNA-recognition domain is localized in the C-terminal half of the AraC protein. Truncated proteins were unable to repress araBAD or araC in vivo, even though they were able to bind all ara operators. We propose that the N-terminal half of AraC is essential for the formation of the DNA loops that are responsible for repression of araBAD and for autoregulation of araC.
Menon
K P
KP
Department of Biological Sciences, University of California, Santa Barbara 93106.
Lee
N L
NL
D051777:AraC Transcription Factor; D001426:Bacterial Proteins; D001483:Base Sequence; D002872:Chromosome Deletion; D004926:Escherichia coli / Q000235:genetics*; D029968:Escherichia coli Proteins; D005809:Genes, Regulator; D005797:Genes, araC; D006706:Homeostasis; D008969:Molecular Sequence Data; D008970:Molecular Weight; D009154:Mutation; D009876:Operon; D011401:Promoter Regions, Genetic; D012097:Repressor Proteins / Q000235:genetics* / Q000302:isolation & purification; D015183:Restriction Mapping; D014157:Transcription Factors / Q000235:genetics*; D014158:Transcription, Genetic; D001616:beta-Galactosidase / Q000235:genetics
D051777:AraC Transcription Factor; C497371:AraC protein, E coli; D001426:Bacterial Proteins; D029968:Escherichia coli Proteins; D012097:Repressor Proteins; D014157:Transcription Factors; D001616:beta-Galactosidase