caa a22 4500 397570058 CHVBK 20180308164838.0 cr unu---uuuuu 161202e199612 xx s 000 0 eng 10.1093/oxfordjournals.jmicro.a023473 doi (NATIONALLICENCE)oxford-10.1093/oxfordjournals.jmicro.a023473 Subcellular Destination of Mutant Peroxisomal Isocitrate Lyase Polypeptides of Candida tropicalis in Saccharomyces cerevisiae [Elektronische Daten] [Naomi Kamasawa, Tomoko Yoshida, Mari Kasahara, Yasushi Kamada, Wen Zou, Mitsuyoshi Ueda, Atsuo Tanaka, Masako Osumi] The peroxisomal isocitrate lyase (ICL) of an n-alkane-utilizable yeast, Candida tropicalis, could be transported into the peroxisomes of Saccharomyces cerevisiae MT8-1 cells grown in an oleic acid medium, even if strains were different. To identify the important regions to direct C. tropicalis ICL (CT-ICL) to its proper subcellular location in S. cerevisiae, we observed the subcellular localization of the expressed products of mutant CT-ICL genes in S. cerevisiae by immunoelectron microscopy. Under the control of the UPR-ICL promoter, which enhanced gene expression by proliferation of the peroxisomes, the following mutant CT-ICL genes were constructed and expressed: (i) truncated or mutant genes in the C-terminal region which apparently contained peroxisomal targeting signals, Δ550, Δ549-550, Δ548-550, Δ548-550 + SKL and Δ340-550; and (ii) four internal deletion mutant genes, Δ41-115, Δ41-239, Δ41-325 and Δ237-339. The results showed that three C-terminal amino acid residues were not essential for targeting the peroxisomes in this enzyme. In cells harboring the deleted internal regions of mutant ICL genes, the expressed polypeptides did not target the peroxisomes and the polypeptides, except for Δ41-115, were mistargeted to mitochondria. We observed an unknown structure which we called a "protein aggregate body (PAB),” in which various mutant CT-ICL polypeptides aggregated. © 1996 Oxford University Press Full-length Papers nationallicence immunoelectron microscopy nationallicence yeast nationallicence isocitrate lyase nationallicence peroxisome (microbody) nationallicence targeting signal nationallicence Kamasawa Naomi Department of Ciemical and Biological Sciences, Faculty of Science, Japan Women's University 2-8-1 Mejiro-dai, Bunkyo-ku, Tokyo, 112 Jupan aut Yoshida Tomoko Department of Ciemical and Biological Sciences, Faculty of Science, Japan Women's University 2-8-1 Mejiro-dai, Bunkyo-ku, Tokyo, 112 Jupan aut Kasahara Mari Department of Ciemical and Biological Sciences, Faculty of Science, Japan Women's University 2-8-1 Mejiro-dai, Bunkyo-ku, Tokyo, 112 Jupan aut Kamada Yasushi Laboratory of Applied Biological Chemistry, Department of Synthetic Chemistry and Biological Chemistry. Graduate School of Engineering, Kyoto University Yoshida, Sakyo-ku, Kyoto, 606-01 Japan aut Zou Wen Laboratory of Applied Biological Chemistry, Department of Synthetic Chemistry and Biological Chemistry. Graduate School of Engineering, Kyoto University Yoshida, Sakyo-ku, Kyoto, 606-01 Japan aut Ueda Mitsuyoshi Laboratory of Applied Biological Chemistry, Department of Synthetic Chemistry and Biological Chemistry. Graduate School of Engineering, Kyoto University Yoshida, Sakyo-ku, Kyoto, 606-01 Japan aut Tanaka Atsuo Laboratory of Applied Biological Chemistry, Department of Synthetic Chemistry and Biological Chemistry. Graduate School of Engineering, Kyoto University Yoshida, Sakyo-ku, Kyoto, 606-01 Japan aut Osumi Masako Department of Ciemical and Biological Sciences, Faculty of Science, Japan Women's University 2-8-1 Mejiro-dai, Bunkyo-ku, Tokyo, 112 Jupan aut Microscopy Oxford University Press 45/6(1996-12), 491-497 0022-0744 45:6<491 1996 45 jmicro https://doi.org/10.1093/oxfordjournals.jmicro.a023473 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1093/oxfordjournals.jmicro.a023473 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Kamasawa Naomi Department of Ciemical and Biological Sciences, Faculty of Science, Japan Women's University 2-8-1 Mejiro-dai, Bunkyo-ku, Tokyo, 112 Jupan aut NATIONALLICENCE 700 1- Yoshida Tomoko Department of Ciemical and Biological Sciences, Faculty of Science, Japan Women's University 2-8-1 Mejiro-dai, Bunkyo-ku, Tokyo, 112 Jupan aut NATIONALLICENCE 700 1- Kasahara Mari Department of Ciemical and Biological Sciences, Faculty of Science, Japan Women's University 2-8-1 Mejiro-dai, Bunkyo-ku, Tokyo, 112 Jupan aut NATIONALLICENCE 700 1- Kamada Yasushi Laboratory of Applied Biological Chemistry, Department of Synthetic Chemistry and Biological Chemistry. Graduate School of Engineering, Kyoto University Yoshida, Sakyo-ku, Kyoto, 606-01 Japan aut NATIONALLICENCE 700 1- Zou Wen Laboratory of Applied Biological Chemistry, Department of Synthetic Chemistry and Biological Chemistry. Graduate School of Engineering, Kyoto University Yoshida, Sakyo-ku, Kyoto, 606-01 Japan aut NATIONALLICENCE 700 1- Ueda Mitsuyoshi Laboratory of Applied Biological Chemistry, Department of Synthetic Chemistry and Biological Chemistry. Graduate School of Engineering, Kyoto University Yoshida, Sakyo-ku, Kyoto, 606-01 Japan aut NATIONALLICENCE 700 1- Tanaka Atsuo Laboratory of Applied Biological Chemistry, Department of Synthetic Chemistry and Biological Chemistry. Graduate School of Engineering, Kyoto University Yoshida, Sakyo-ku, Kyoto, 606-01 Japan aut NATIONALLICENCE 700 1- Osumi Masako Department of Ciemical and Biological Sciences, Faculty of Science, Japan Women's University 2-8-1 Mejiro-dai, Bunkyo-ku, Tokyo, 112 Jupan aut NATIONALLICENCE 773 0- Microscopy Oxford University Press 45/6(1996-12), 491-497 0022-0744 45:6<491 1996 45 jmicro Metadata rights reserved CC BY-NC-4.0 http://creativecommons.org/licenses/by-nc/4.0 nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-oxford