Structural specificity of polyamines in modulating the binding of estrogen receptor to potential Z-DNA forming sequences.
Estrogen receptor (ER) is a gene-regulatory protein that mediates the action of estradiol. In order to examine the role of conformational dynamics of DNA in estrogenic regulation of gene expression, we studied the binding of ER to poly(dA-dC).poly(dG-dT) which undergoes transition to a left-handed Z-DNA form. This type of dinucleotide repeats are widely distributed in mammalian genome and are present in estrogen response elements. Binding affinity of ER for the polynucleotide was assessed by its ability to release ER bound to DNA-cellulose. ER binding by poly(dA-dC).poly(dG-dT) was enhanced in the presence of an endogenous polyamine, spermidine, H2N(CH2)4NH(CH2)3NH2. The concentration of spermidine required for facilitating 50% elution of ER (EC50) was 75 microM. This EC50 increased to 500 microM for a spermidine homolog, H2N(CH2)8NH(CH2)3NH2, demonstrating polyamine structural specificity. Spectroscopic measurements showed that the presence of 100-200 microM spermidine initiated changes in the conformation of the polynucleotide indicative of Z-DNA form, but a major alteration to Z-DNA spectrum occurred only at 300 microM concentration. These data suggest that ER favors DNA sequences poised for Z-DNA transition. The efficacy of spermidine homologs in facilitating ER-DNA interaction may be important in predicting their efficiency to replace cellular functions of spermidine.
Thomas
T
T
Department of Environmental and Community Medicine, University of Medicine and Dentistry of New Jersey-Robert Wood Johnson Medical School, Piscataway 08854.
Thomas
T J
TJ
D000818:Animals; D001483:Base Sequence; D002942:Circular Dichroism; D004247:DNA / Q000737:chemistry* / Q000378:metabolism; D008969:Molecular Sequence Data; D009690:Nucleic Acid Conformation; D011089:Polydeoxyribonucleotides / Q000378:metabolism; D011817:Rabbits; D011960:Receptors, Estrogen / Q000378:metabolism*; D012965:Sodium Chloride / Q000494:pharmacology; D013095:Spermidine / Q000737:chemistry / Q000494:pharmacology*; D013329:Structure-Activity Relationship
D011089:Polydeoxyribonucleotides; D011960:Receptors, Estrogen; D012965:Sodium Chloride; C033927:poly(dA-dC).poly(dG-dT); D004247:DNA; D013095:Spermidine