Abstract: A chimeric transactivator comprises a transcription activation domain, a repressor protein DNA binding domain and the bacterial DNA gyrase B subunit. A target gene is operatively linked to operator DNA sequences recognized by the repressor binding domain. The addition of the antibiotic coumermycin results in a coumermycin-switched dimerization of the transactivator, which then binds to operator DNA sequences and activates transcription of the target gene. The addition of novobiocin switches off expression of the target gene by abolishing coumermycin-induced dimerization of the transactivator.

Abstract: A chimeric transactivator comprises a transcription activation domain, a repressor protein DNA binding domain and the bacterial DNA gyrase B subunit. A target gene is operatively linked to operator DNA sequences recognized by the repressor binding domain. The addition of the antibiotic coumermycin results in a coumermycin-switched dimerization of the transactivator, which then binds to operator DNA sequences and activates transcription of the target gene. The addition of novobiocin switches off expression of the target gene by abolishing coumermycin-induced dimerization of the transactivator.

Abstract: A chimeric transactivator comprises a transcription activation domain, a repressor protein DNA binding domain and the bacterial DNA gyrase B subunit. A target gene is operatively linked to operator DNA sequences recognized by the repressor binding domain. The addition of the antibiotic coumermycin results in a coumermycin-switched dimerization of the transactivator, which then binds to operator DNA sequences and activates transcription of the target gene. The addition of novobiocin switches off expression of the target gene by abolishing coumermycin-induced dimerization of the transactivator.

Abstract: A chimeric transactivator comprises a transcription activation domain, a repressor protein DNA binding domain and the bacterial DNA gyrase B subunit. A target gene is operatively linked to operator DNA sequences recognized by the repressor binding domain. The addition of the antibiotic coumermycin results in a coumermycin-switched dimerization of the transactivator, which then binds to operator DNA sequences and activates transcription of the target gene. The addition of novobiocin switches off expression of the target gene by abolishing coumermycin-induced dimerization of the transactivator.

Abstract: The present invention provides compositions, methods, and kits for identifying agents that are capable of disrupting protein-protein interaction in a mammalian reverse two-hybrid system. A tetracycline repressor protein was fused with the inhibitory KRAB domain as a suppressor to tightly regulate the reverse two-hybrid system for mammalian cells. Binding of the chimeric protein to the tetracycline operator sequence provided within a promoter entirely suppressed the expression of two interactive proteins. When both an inducer and a candidate protein-protein interaction disrupting agent such as a drug were added in the cell culture together, the reporter gene was either activated by the interaction of protein-pair if the drug was ineffective or remained silent due to the disruption of the protein-protein interaction by the effective drug. The plasmids for the suppressor and the reporter were integrated into chromosomes.

Abstract: The invention features methods of treating insulin-induced obesity, weight gain, and other conditions associated with hyperinsulinemia.