title: Analysis of heterodimer formation by the Escherichia coli trp repressor.
abstract: The trp repressor of Escherichia coli is a dimeric DNA-binding protein that regulates transcription of several operons concerned with tryptophan metabolism. Although heterodimer formation between mutant and wild type subunits occurs readily in vivo, comparable heterodimers could be formed in vitro only under extreme conditions. To explain this difference we analyzed trp repressor dimer formation and dissociation using an in vitro transcription/translation system. Nascent wild type or mutant repressor polypeptides, synthesized in the presence of an excess of a second repressor, were invariably incorporated into heterodimers. In contrast, previously synthesized and assembled wild type dimers appeared to be refractory to dissociation, since they did not form heterodimers. However, previously synthesized mutant dimeric repressors that were defective in tryptophan binding readily dissociated and formed heterodimers. We noted that the ability of a dimeric repressor to dissociate under our conditions correlated inversely with its affinity for tryptophan. Consistent with this conclusion, we found that dissociation of the wild type aporepressor (no added tryptophan) was appreciably more rapid than dissociation of the tryptophan-saturated wild type repressor.
authors.affiliation: Department of Biological Sciences, Stanford University, California 94305.
authors.forename: B K
authors.initials: BK
authors.lastname: Hurlburt
authors.forename: C
authors.initials: C
authors.lastname: Yanofsky
mesh_terms: D001426:Bacterial Proteins; D001665:Binding Sites; D001704:Biopolymers; D004926:Escherichia coli / Q000378:metabolism*; D014176:Protein Biosynthesis; D012097:Repressor Proteins / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism*; D014158:Transcription, Genetic; D014364:Tryptophan / Q000378:metabolism; D029968:Escherichia coli Proteins
chemical_list: D001426:Bacterial Proteins; D001704:Biopolymers; D012097:Repressor Proteins; D014364:Tryptophan; C027190:TRPR protein, E coli; D029968:Escherichia coli Proteins