Individual Expression of Candida tropicalis Peroxisomal and Mitochondrial Carnitine Acetyltransferase-Encoding Genes and Subcellular Localization of the Products in Saccharomyces cerevisiae.
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概要
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In an n-alkane-assimilating yeast, Candida tropicalis, carnitine acetyltransferase (CAT; EC 2. 3. 1. 7) was localized in both peroxisomes and mitochondria. Both CATs were encoded by one gene, CT-CAT, although the initiation sites of translation were suggested to be different. In the present study, the genes corresponding to the supposed C. tropicalis peroxisomal and mitochondrial CATs, which were truncated from the CT-CAT gene, were individually expressed in Saccharomyces cerevisiae, using the C. tropicalis isocitrate lyase promoter (UPR-ICL), which is inducible by oleic acid in concert with proliferation of peroxisomes in S. cerevisiae [Umemura, K., Atomi, H., Kanai, T., Teranishi, Y., Ueda, M., and Tanaka, A. (1995) Appl. Microbiol. Biotechnol. 43, 489-492]. The 71 kDa precursor of mitochondrial CAT, initiating at the first Met, was found to be processed to the mature size (66 kDa) in S. cerevisiae and immunoelectronmicroscopical observation revealed that this enzyme was localized in mitochondria. On the other hand, 68 kDa CAT, initiating at the second Met (residue No. 19), had no cleavable signal and was translocated into peroxisomes and cytosol, but not into mitochondria. The amino-terminal amino acid sequences of individually expressed CATs were identical to those of CATs isolated from alkane-grown C. tropicalis cells, respectively. These results demonstrated that only the 71 kDa protein yielded the 66 kDa protein and that peroxisomal and mitochondria) CATs arose from the difference in the initiation sites of translation.
- 社団法人 日本生化学会の論文
著者
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Atomi Haruyuki
Laboratory Of Applied Biological Chemistry Department Of Synthetic Chemistry And Biological Chemistr
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KAWACHI Hiroyuki
Laboratory of Applied biological Chemistry, Department of Synthetic Chemistry and Biological Chemist
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Kobayashi Keiko
Department Of Biochemical Pharmacology Faculty Of Pharmaceutical Sciences Chiba University
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Kamasawa Naomi
Department Of Chemical And Biological Sciences Faculty Of Science Japan Women's University
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Tanaka Atsuo
Laboratory Of Applied Biological Chemistry Department Of Synthetic Chemistry And Biological Chamistr
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Ueda Mitsuyoshi
Laboratory Of Applied Biological Chemistry Department Of Synthetic Chemistry And Biological Chemistr
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Osumi Masako
Department Of Biology Faculty Of Science Osaka City University
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Yoshida Tomoko
Department of Biochemical Toxicology, School of Pharmacy Showa University
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HASHIMOTO Noriko
Department of Allergy and Immunology, National Research Institute for Child Health and Development
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OSUMI Masako
Department of Chemical and Biological Sciences, Faculty of Science, Japan Women's University
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Ueda Mitsuyoshi
Laboratory of Applied Biological Chemistry, Department of Synthetic Chemistry and Biological Chemistry, Graduate School of Engineering, Kyoto University
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