Effect of redox conditions on the DNA-binding efficiency of the retinoic acid receptor zinc-finger.
Retinoic acid and its derivatives are involved in many important biological processes. In the present study, we have shown that the DNA binding domain of the retinoic acid receptor, which contains two zinc fingers with the Zn(II) tetrahedrally coordinated by four Cys, is susceptible to intracellularly relevant oxidizing agents. In the presence of hydrogen peroxide or hypochlorite, the zinc-finger DNA binding activity was abolished in a concentration dependent manner. The loss of DNA binding activity was correlated with the release of Zn(II) from the zinc-finger motif as a consequence of Zn(II)-thiolate bond oxidation. A combination of glutathione and Zn(II) was able to restore the activity, suggesting that oxidation of the zinc-finger by hydrogen peroxide or hypochlorite resulted in the formation of disulfide bonds between the Cys present in the Zn(II)-binding motif. Our results indicate that in situations of oxidative-stress zinc-finger containing transcription factors may be particularly susceptible to oxidation, resulting in the disruption of control and regulation of gene expression.
Casadevall
M
M
Department of Biochemistry Research, Hospital for Sick Children, Toronto, Ont., Canada.
Sarkar
B
B
D000818:Animals; D001483:Base Sequence; D001665:Binding Sites / Q000235:genetics; D004247:DNA / Q000235:genetics / Q000378:metabolism*; D004268:DNA-Binding Proteins / Q000737:chemistry* / Q000378:metabolism*; D005786:Gene Expression Regulation; D066298:In Vitro Techniques; D051379:Mice; D010084:Oxidation-Reduction; D018384:Oxidative Stress; D018168:Receptors, Retinoic Acid / Q000737:chemistry* / Q000378:metabolism*; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism; D014157:Transcription Factors / Q000737:chemistry / Q000378:metabolism; D015032:Zinc / Q000378:metabolism; D016335:Zinc Fingers
D004268:DNA-Binding Proteins; D018168:Receptors, Retinoic Acid; D011994:Recombinant Proteins; D014157:Transcription Factors; D004247:DNA; D015032:Zinc