Specific interactions by the N-terminal arm inhibit self-association of the AraC dimerization domain.
Deletion of the regulatory N-terminal arms of the AraC protein from its dimerization domain fragments increases the susceptibility of the dimerization domain to form a series of higher order polymers by indefinite self-association. We investigated how the normal presence of the arm inhibits this self-association. One possibility is that arms can act as an entropic bristles to interfere with the approach of other macromolecules, thereby decreasing collision frequencies. We examined the repulsive effect of flexible arms by measuring the rate of trypsin cleavage of a specially constructed ubiquitin-arm protein. Adding an arm to ubiquitin or increasing its length produced only a modest repulsive effect. This suggests that arms such as the N-terminal arm of AraC do not reduce self-association by entropic exclusion. We consequently tested the hypothesis that the arm on AraC reduces self-association by binding to the core of the dimerization domain even in the absence of arabinose. The behaviors of dimerization domain mutants containing deletions or alterations in the N-terminal arms substantiate this hypothesis. Apparently, interactions between the N-terminal arm and the dimerization domain core position the arm to interfere with the protein-protein contacts necessary for self-association.
Weldon
John E
JE
Department of Biology, Johns Hopkins University, Baltimore, Maryland 21218, USA.
Schleif
Robert F
RF
D051777:AraC Transcription Factor / Q000737:chemistry* / Q000378:metabolism*; D001665:Binding Sites; D019281:Dimerization; D019277:Entropy; D004926:Escherichia coli; D008954:Models, Biological; D050505:Mutant Proteins / Q000737:chemistry / Q000378:metabolism; D009154:Mutation / Q000502:physiology; D011485:Protein Binding; D015202:Protein Engineering / Q000379:methods; D017510:Protein Folding; D017434:Protein Structure, Tertiary / Q000502:physiology; D012995:Solubility; D014357:Trypsin / Q000378:metabolism
D051777:AraC Transcription Factor; D050505:Mutant Proteins; D014357:Trypsin