Synergistic activation of regulatory elements by rel proteins and a steroid receptor

Abstract

Interaction of Rel proteins and steroid receptors is known to result in repression of target genes. Here we describe the discovery of a new mechanism in which Rel proteins and steroid receptors act synergistically to activate a regulatory element. This mechanism is shown to influence the expression of the brain-specific 5HT1A receptor wherein the estrogen receptor acts synergistically with the Nuclear Factor kappa B to enhance the activity of the promoter for the 5HT1A receptor gene. In addition, synergistic effects of Rel proteins with the mineralocorticoid receptor were observed, showing that synergism with Rel proteins may be expected for other steroid receptors as well. The synergism between Rel proteins and estrogen receptor or mineralocorticoid receptor provides a tool for the development of compounds that interact with the estrogen or mineralocorticoid receptor in such a way that only the synergistic effect is modulated whereas other effects are left intact. Such compounds would be invaluable for the development of medicaments for the treatment of diseases of the central nervous or cardiovascular system and other disorders in which Rel proteins play a role.

1 - 20 . (canceled)  
     
     
         21 . A method for the identification of compounds capable of modulating a synergistic effect of a steroid receptor and a Rel protein on regulatory elements, comprising the steps of: 
 providing a cell comprising a regulatory element capable of being synergistically activated by a steroid receptor and a Rel protein, said cell additionally comprising sufficient levels of said steroid receptor and said Rel protein to allow synergistic activation of the regulatory element;    contacting said cell with at least one compound; and,    determining whether the activation of the regulatory element is modulated by the compound, wherein the regulatory element is a promoter for the E-selectin gene.    
     
     
         22 . The method according to  claim 21 , wherein said steroid receptor is selected from the group consisting of the estrogen receptor α, the estrogen receptor β, and the nineralocorticoid receptor.  
     
     
         23 . The method according to  claim 21 , wherein the Rel protein is selected from the group consisting of NF-κB1, Lyt-10, cRel, RelA and RelB.  
     
     
         24 . A cell transfected with a nucleic acid comprising a promotor for the E-selectin gene and a reporter gene capable of signaling the activation of the promoter for the E-selectin gene, said cell further comprising; 
 a nucleic acid encoding a steroid receptor and capable of functionally expressing said steroid receptor within said cell, and    a nucleic acid encoding a Rel protein and capable of functionally expressing said Rel protein within said cell.    
     
     
         25 . The cell according to  claim 24 , wherein said steroid receptor is selected from the group consisting of the estrogen receptor a, the estrogen receptor 6 and the mineralocorticoid receptor.  
     
     
         26 . The cell according to  claim 24 , wherein the Rel protein is selected from the group consisting of NF-κB1, Lyt-10, cRel, RelA and RelB.