Genome-wide drug-induced haploinsufficient screening of fission yeast for 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 genomewide 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 microM. The initial screening of 4,158 mutants in 384-well plates using robotics was performed at concentrations of 2, 3, and 4 microM. 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 acetyl transferase. The target mutants showed 65% growth inhibition in response to HC compared with wild-type controls, as shown by liquid culture assay.
Baek
Seung Tae
ST
Functional Genomics Research Center, Korea Research Institute of Bioscience and Biotechnology , Daejeon 305-333, Korea.
Kim
Dong-Uk
DU
Han
Sangjo
S
Woo
Im Sun
IS
Nam
Miyoung
M
Kim
Lila
L
Heo
Kyung-Sun
KS
Lee
Hyemi
H
Hwang
Hye-Rim
HR
Choi
Shin-Jung
SJ
Won
Misun
M
Lee
Minho
M
Park
Song-Kyu
SK
Lee
Sunghou
S
Kwon
Ho-Jung
HJ
Maeng
Pil Jae
PJ
Park
Hee-Moon
HM
Park
Youngwoo
Y
Kim
Dongsup
D
Hoe
Kwang-Lae
KL
D003474:Curcumin / Q000031:analogs & derivatives* / Q000494:pharmacology; D004353:Drug Evaluation, Preclinical; D017353:Gene Deletion; D016681:Genome, Fungal; D006238:Haploidy; D006579:Heterozygote; D006834:Hydrazines / Q000494:pharmacology*; D020128:Inhibitory Concentration 50; D012568:Schizosaccharomyces / Q000187:drug effects* / Q000235:genetics / Q000254:growth & development / Q000378:metabolism; D029702:Schizosaccharomyces pombe Proteins / Q000037:antagonists & inhibitors / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism
D006834:Hydrazines; D029702:Schizosaccharomyces pombe Proteins; C475807:hydrazinocurcumin; D003474:Curcumin