Genetic Analysis of Growth Inhibition of Yeast Cells Caused by Expression of Aspergillus oryzae RNase T1
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概要
- 論文の詳細を見る
Even though most fungal hydrolytic enzymes have been successfully secreted in S. cerevisiae cells by expression of corresponding cDNA, overexpression of A. oryzae RNase T1 causes severe growth inhibition in yeast. We observed that yeast strains carrying RNase T1 cDNA under control of the GAL1 promoter with a single-copy vector were able to grow on galactose medium while those with a multi-copy vector were not. It was found that overexpression of three mutated versions of RNase T1 with low enzymatic activity did not affect the growth. We also observed that expression of RNase T1 without a signal sequence severely inhibited growth of the transformant even on the single-copy plasmid. Subcellular fractionation showed that overexpressed myctagged RNase T1 was localized in the membrane fraction. In the yeast secretory pathway, while the mutants defective in translocation into the ER, ER-Golgi trafficking and vacuole formation had severe growth inhibition during expression of RNase T1 from the singlecopy plasmid. These results suggest that a mislocalization of active RNase T1 in cytosol by overflow from the secretory apparatus has toxic effects on the host cells.
- 社団法人日本農芸化学会の論文
- 2000-10-23
著者
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Kitamoto Katsuhiko
Department of Biotechnology, The University of Tokyo
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NAKAJIMA Harushi
Department of Biotechnology, The University of Tokyo
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Nonaka Gen
Usaien Pharmaceutical Co. Ltd.
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Nonaka Gen
Procter And Gamble Far East Inc.
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Kitamoto Katsuhiko
Department Of Applied Biological Chemistry And Department Of Applied Biological Engineering Graduate
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NONAKA Gen
Department of Biotechnology, The University of Tokyo
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ISHIKAWA Tomoki
Department of Biotechnology, The University of Tokyo
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LIU Tintin
Department of Biotechnology, The University of Tokyo
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Nonaka Gen
Department Of Biotechnology The University Of Tokyo
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Nakajima Harushi
Department Of Agricultural Chemistry The University Of Tokyo
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Nakajima Harushi
Department Of Biotechnology The University Of Tokyo
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Ishikawa Tomoki
Department Of Biotechnology The University Of Tokyo
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Liu Tintin
Department Of Biotechnology The University Of Tokyo
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Liu Tin-tin
Department of Biotechnology, The University of Tokyo
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