Genome-wide Drug-induced Haploinsufficiency Screening of Fission Yeastfor Identification of Hydrazinocurcumin Targets

Hydrazinocurcumin (HC), a synthetic derivative of curcumin, has been reported to inhibit angiogenesis via unknown mechanisms. Understanding the molecular mechanisms of the drug’s action is important for the development of improved compounds with better pharmacological properties. A genome-wide drug-induced haploinsufficiency screening of fission yeast gene deletion mutants has been applied to identify drug targets of HC. As a first step, the 50% inhibition concentration (IC50) of HC was determined to be 2.2 μM. The initial screening of 4,158 mutants in 384-well plates using robotics was performed at concentrations of 2, 3, and 4 μM. A second screening was performed to detect sensitivity to HC on the plates. The first screening revealed 178 candidates, and the second screening resulted in 13 candidates, following the elimination of 165 false positives. Final filtering of the condition-dependent haploinsufficient genes gave eight target genes. Analysis of the specific targets of HC has shown that they are related to septum formation and the general transcription processes, which may be related to histone acetyltransferase. The target mutants showed 65% growth inhibition in response to HC compared with wild-type controls, as shown by liquid culture assay. Hydrazinocurcumin (HC), a synthetic derivative of curcumin,has been reported to inhibit angiogenesis via unknownmechanisms. Understanding the molecular mechanismsof the drug’s action is important for the development ofimproved compounds with better pharmacological properties. A genome-wide drug-induced haploinsufficiency screeningof fission yeast gene deletion mutants has been applied toidentify drug targets of HC. As a first step, the 50%inhibition concentration (IC50) of HC was determined tobe 2.2 μM. The initial screening of 4,158 mutants in 384-wellplates using robotics was performed at concentrations of2, 3, and 4 μM. A second screening was performed to detectsensitivity to HC on the plates. The first screening revealed178 candidates, and the second screening resulted in 13candidates, following the elimination of 165 false positives. Final filtering of the condition-dependent haploinsufficientgenes gave eight target genes. Analysis of the specific targetsof HC has shown that they are related to septum formationand the general transcription processes, which may berelated to histone acetyltransferase. The target mutantsshowed 65% growth inhibition in response to HC comparedwith wild-type controls, as show