Development of novel anti- @entity5 drug leads that target regulators of protein homeostasis is a formidable task in modern pharmacology. Finding specific inhibitors of @entity1 @entity1457 Factor 1 ( @entity22585 ) has proven to be a challenging task, while screening for inhibitors of @entity1 @entity1605 ( @entity42782 ) has never been described. We report the development of a novel system based on an in vivo cell growth restoration assay designed to identify specific inhibitors of @entity1 @entity42782 in a high-throughput format. This system utilizes a humanized @entity375 in which the master regulator of molecular chaperone genes, @entity375 HSF, has been replaced with @entity42782 with no detrimental effect on cell growth. This replacement preserves the general regulatory patterns of genes encoding major molecular chaperones including @entity2285 and @entity4041 . The controlled overexpression of @entity42782 creates a slow-growth phenotype, which is the basis of the growth restoration assay used for high-throughput screening. The phenotype is most robust when cells are cultured at 25   C, while incubation at temperatures greater than 30   C leads to compensation of the phenotype. Overexpression of @entity42782 causes overexpression of molecular chaperones which is a likely cause of the slowed growth. Our assay is characterized by two unique advantages. First, screening takes place in physiologically relevant, in vivo conditions. Second, hits in our screen will be of medically relevant potency, as compounds that completely inhibit @entity42782 function will further inhibit cell growth and therefore will not be scored as hits. This caveat biases our screening system for compounds capable of restoring @entity42782 activity to a physiologically normal level without completely inhibiting this essential system.

High-throughput screening system for inhibitors of XXXX @entity1605 .

@entity1 :: ('9606', 'Species') :: ['human'] @entity1605 :: ('OMIM:217000', 'Disease') :: ['Heat Shock Factor 2'] @entity42782 :: ('3298', 'Gene') :: ['HSF2', 'hHSF2'] @entity4041 :: ('3320', 'Gene') :: ['Hsp90'] @entity2285 :: ('3308', 'Gene') :: ['Hsp70'] @entity5 :: ('MESH:D009369', 'Disease') :: ['cancer'] @entity1457 :: ('MESH:D012769', 'Disease') :: ['Heat Shock'] @entity22585 :: ('3297', 'Gene') :: ['hHSF1'] @entity375 :: ('4932', 'Species') :: ['yeast strain', 'yeast']
@entity1 :: (9606,Species) :: ['human']
