A cell-based assay for identification of small chemical compounds that inhibit Supt4h/Supt5h protein complex formation
碩士 === 國立陽明大學 === 生化暨分子生物研究所 === 102 === Huntington’s disease (HD) is an inherited neurological disorder caused by abnormal expansion of CAG repeats, which encode a long stretch of polyglutamine, in Huntingtin (HTT) gene. While the expression of mutant HTT contributes a gain-of-function for pathogen...
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ndltd-TW-102YM0051070392015-10-13T23:50:23Z http://ndltd.ncl.edu.tw/handle/55170386578516965082 A cell-based assay for identification of small chemical compounds that inhibit Supt4h/Supt5h protein complex formation 篩選Supt4h/Supt5h蛋白質聚合體之抑制劑 Yun-Yun Wu 吳畇芸 碩士 國立陽明大學 生化暨分子生物研究所 102 Huntington’s disease (HD) is an inherited neurological disorder caused by abnormal expansion of CAG repeats, which encode a long stretch of polyglutamine, in Huntingtin (HTT) gene. While the expression of mutant HTT contributes a gain-of-function for pathogenesis of HD, the biological activity of normal HTT is essential for neuron development and survival. For this reason, it is an optimal goal having HD treatments that inhibit the expression of HTT alleles in a selective manner. Currently, effective treatment is not available for this disorder and the development of therapeutic compounds is urgently needed. In our previous study, we demonstrated that Supt4h down-regulation can selectively reduce mutant HTT gene expression without an overt impact on the expression of wild-type HTT. Furthermore, the function of Supt4h in supporting RNA polymerase II transcribing through CAG expansion region is dependent on the formation of Supt4h/5h complex. Here, based on Bimolecular Fluorescence Complementation (BiFC), we established a cell-based assay system to monitor the interaction between Supt4h and Supt5h, and also applied this system to identify small chemical compounds that could interfere with the Supt4h/Supt5h complex formation. Among 60,000 compounds screened, 7 hits were identified with an inhibitory effect on the fluorescence signal of BiFC. These compounds were subjected to further characterization to assure their activities on Supt4h/5h protein-protein interaction. Tzu-Hao Cheng 鄭子豪 2014 學位論文 ; thesis 54 zh-TW |
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碩士 === 國立陽明大學 === 生化暨分子生物研究所 === 102 === Huntington’s disease (HD) is an inherited neurological disorder caused by abnormal expansion of CAG repeats, which encode a long stretch of polyglutamine, in Huntingtin (HTT) gene. While the expression of mutant HTT contributes a gain-of-function for pathogenesis of HD, the biological activity of normal HTT is essential for neuron development and survival. For this reason, it is an optimal goal having HD treatments that inhibit the expression of HTT alleles in a selective manner. Currently, effective treatment is not available for this disorder and the development of therapeutic compounds is urgently needed.
In our previous study, we demonstrated that Supt4h down-regulation can selectively reduce mutant HTT gene expression without an overt impact on the expression of wild-type HTT. Furthermore, the function of Supt4h in supporting RNA polymerase II transcribing through CAG expansion region is dependent on the formation of Supt4h/5h complex.
Here, based on Bimolecular Fluorescence Complementation (BiFC), we established a cell-based assay system to monitor the interaction between Supt4h and Supt5h, and also applied this system to identify small chemical compounds that could interfere with the Supt4h/Supt5h complex formation. Among 60,000 compounds screened, 7 hits were identified with an inhibitory effect on the fluorescence signal of BiFC. These compounds were subjected to further characterization to assure their activities on Supt4h/5h protein-protein interaction.
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author2 |
Tzu-Hao Cheng |
author_facet |
Tzu-Hao Cheng Yun-Yun Wu 吳畇芸 |
author |
Yun-Yun Wu 吳畇芸 |
spellingShingle |
Yun-Yun Wu 吳畇芸 A cell-based assay for identification of small chemical compounds that inhibit Supt4h/Supt5h protein complex formation |
author_sort |
Yun-Yun Wu |
title |
A cell-based assay for identification of small chemical compounds that inhibit Supt4h/Supt5h protein complex formation |
title_short |
A cell-based assay for identification of small chemical compounds that inhibit Supt4h/Supt5h protein complex formation |
title_full |
A cell-based assay for identification of small chemical compounds that inhibit Supt4h/Supt5h protein complex formation |
title_fullStr |
A cell-based assay for identification of small chemical compounds that inhibit Supt4h/Supt5h protein complex formation |
title_full_unstemmed |
A cell-based assay for identification of small chemical compounds that inhibit Supt4h/Supt5h protein complex formation |
title_sort |
cell-based assay for identification of small chemical compounds that inhibit supt4h/supt5h protein complex formation |
publishDate |
2014 |
url |
http://ndltd.ncl.edu.tw/handle/55170386578516965082 |
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