High throughput screens for small-molecule inhibitors of the nuclear receptor-like pathway regulating multidrug resistance in fungi
The present invention relates to the identification of molecular mechanisms associated with multidrug resistance (MDR) in fungal infections. More specifically, fungi harbor a nuclear receptor-like pathway controlling MDR, which represents a novel therapeutic target for the treatment of MDR in pathogenic fungi such as C. glabrata.
1. A method for the high throughput screening for agents that inhibit the multidrug resistance pathway in fungi comprising:
providing a solution of fluorescein-labeled Pdr1p activation domain, a Gal11p
KIX domain and a test agent; and
comparing the fluorescence polarization of the solution of fluorescein-labeled Pdr1p activation domain, the Gal11p KIX domain and the test agent, with the fluorescence polarization of a solution of fluorescein-labeled Pdr1p activation domain and a Gal11p KIX domain;
wherein the fluorescein-labeled Pdr1p activation domain consists of fluorescein and 30 amino acids of the CgPdr1p activation domain, having the amino acid sequence
LGTLDEFVNKGDLNELYNSLWGDLFSDVYL.
(SEQ ID NO: 14)
2. The method of claim 1 , wherein the Gal11p-KIX domain is CgGal11Ap KIX.
3. The method of claim 1 , wherein the Gal11p-KIX domain is GST-tagged.
4. The method of claim 1 , wherein the comparing is automated.