Radiation abolishes inducer binding to lactose repressor.
The lactose operon functions under the control of the repressor-operator system. Binding of the repressor to the operator prevents the expression of the structural genes. This interaction can be destroyed by the binding of an inducer to the repressor. If ionizing radiations damage the partners, a dramatic dysfunction of the regulation system may be expected. We showed previously that gamma irradiation hinders repressor-operator binding through protein damage. Here we show that irradiation of the repressor abolishes the binding of the gratuitous inducer isopropyl-1-beta-D-thiogalactoside (IPTG) to the repressor. The observed lack of release of the repressor from the complex results from the loss of the ability of the inducer to bind to the repressor due to the destruction of the IPTG binding site. Fluorescence measurements show that both tryptophan residues located in or near the IPTG binding site are damaged. Since tryptophan damage is strongly correlated with the loss of IPTG binding ability, we conclude that it plays a critical role in the effect. A model was built that takes into account the kinetic analysis of damage production and the observed protection of its binding site by IPTG. This model satisfactorily accounts for the experimental results and allows us to understand the radiation-induced effects.
Gillard
Nathalie
N
Centre de Biophysique Moléculaire, CNRS, F-45071 Orléans Cedex 2, France.
Spotheim-Maurizot
Mélanie
M
Charlier
Michel
M
D001426:Bacterial Proteins / Q000737:chemistry* / Q000528:radiation effects*; D001665:Binding Sites / Q000528:radiation effects; D003198:Computer Simulation; D004307:Dose-Response Relationship, Radiation; D005720:Gamma Rays; D007544:Isopropyl Thiogalactoside / Q000737:chemistry* / Q000528:radiation effects*; D056968:Lac Repressors; D008956:Models, Chemical; D011485:Protein Binding / Q000528:radiation effects; D011829:Radiation Dosage; D012097:Repressor Proteins / Q000737:chemistry* / Q000528:radiation effects*; D014364:Tryptophan / Q000737:chemistry / Q000528:radiation effects
D001426:Bacterial Proteins; D056968:Lac Repressors; D012097:Repressor Proteins; D007544:Isopropyl Thiogalactoside; D014364:Tryptophan