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J. Microbiol. Biotechnol. 2008; 18(2): 263-269

Published online February 28, 2008

Copyright © The Korean Society for Microbiology and Biotechnology.

Genome-wide Drug-induced Haploinsufficient Screening of Fission Yeast for Identification of Hydrazinocurcumin Targets

Dongsup Kim , Kwang-Lae Hoe 1*, Baek, Seung Tae 1*, Dong-Uk Kim 1*, Sangjo Han 1*, Im Sun Woo 1*, Miyoung Nam 1*, Lila Kim 1*, Kyung-Sun Heo 1*, Hyemi Lee 1*, Hye-Rim Hwang 1*, Shin-Jung Choi 1*, Misun Won 1*, Minho Lee 1*, Song-Kyu Park 2*, Sunghou Lee 3*, Ho-Jung Kwon 4*, Pil Jae Maeng 5*, Hee-Moon Park 5* and Youngwoo Park 6*

Department of Bio and Brain Engineering, Korea Advanced Institute of Science and Technology (KAIST), Daejeon 305-701, Korea, 1Functional Genomics Research Center, Korea Research Institute of Bioscience and Biotechnology (KRIBB), Daejeon 305-333, Korea, 2Bio-Evaluation Center, Korea Research Institute of Bioscience and Biotechnology (KRIBB), Daejeon 305-333, Korea, 3Department of Biotechnology and Informatics, Sangmyung University, Cheonan 330-720,korea, 4Department of Biology, Yonsei University, Seoul 120-749, Korea, 5Department of Microbiology, School of Biological Science and Biotechnology, Chungnam National University, Daejeon 305-764, Korea, 6Therapeutic Antibody Research Center, Korea Research Institute of Bioscience and Biotechnology (KRIBB), Daejeon 305-333, Korea

Abstract

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 $(IC_{50})$ of HC was determined to be $2.2{\mu}M$. The initial screening of 4,158 mutants in 384-well plates using robotics was performed at concentrations of 2, 3, and $4{\mu}M$. 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.

Keywords: Haploinsufficiency, hydrazinocurcumin, septin, taf4, yeast