Peroxisome targeting signal type 1 (PTS1) receptor is involved in import of both PTS1 and PTS2: Studies with PEX5-defective CHO cell mutants

Hidenori Otera, Kanji Okumoto, Keita Tateishi, Yuka Ikoma, Eiko Matsuda, Maki Nishimura, Toshiro Tsukamoto, Takashi Osumi, Kazumasa Ohashi, Osamu Higuchi, Yukio Fujiki

研究成果: ジャーナルへの寄稿記事

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抄録

To investigate the mechanisms of peroxisome assembly and the molecular basis of peroxisome assembly disorders, we isolated and characterized a peroxisome-deficient CHO cell mutant, ZP139, which was found to belong to human complementation group II, the same group as that of our earlier mutant, ZP105. These mutants had a phenotypic deficiency in the import of peroxisomal targeting signal type 1 (PTS1) proteins. Amino-terminal extension signal (PTS2)-mediated transport, including that of 3-ketoacyl coenzyme A thiolase, was also defective in ZP105 but not in ZP139. PEX5 cDNA, encoding the PTS1 receptor (PTS1R), was isolated from wild-type CHO-K1 cells. PTS1R's deduced primary sequence comprised 595 amino acids, 7 amino acids less than the human homolog, and contained seven tetratricopeptide repeat (TPR) motifs at the C- terminal region. Chinese hamster PTS1R showed 94, 28, and 24% amino acid identity with PTS1Rs from humans, Pichia pastoris, and Saccharomyces cerevisiae, respectively. A PTS1R isoform (PTS1RL) with 632 amino acid residues was identified in CHO cells; for PTSIR, 37 amino acids were inserted between residues at positions 215 and 216 of a shorter isoform (PTS1RS). Southern blot analysis of CHO cell genomic DNA suggested that these two isoforms are derived from a single gene. Both types of PEX5 complemented impaired import of PTS1 in mutants ZP105 and ZP139, PTS2 import in ZP105 was rescued only by PTS1RL. This finding strongly suggests that PTS1RL is also involved in the transport of PTS2. Mutations in PEX5 were determined by reverse transcription-PCR: a G-to-A transition resulted in one amino acid substitution: Gly298Glu of PTS1RS (G335E of PTS1RL) in ZP105 and Gly485Glu of PTS1RS (G522E of PTS1RL) in ZP139. Both mutations were in the TPR domains (TPR1 and TPR6), suggesting the functional consequence of these domains in protein translocation. The implications of these mutations are discussed.

元の言語英語
ページ(範囲)388-399
ページ数12
ジャーナルMolecular and cellular biology
18
発行部数1
出版物ステータス出版済み - 1 1 1998

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Peroxisomes
CHO Cells
Amino Acids
Protein Isoforms
Mutation
Pichia
Protein Transport
Amino Acid Substitution
Coenzyme A
Southern Blotting
Cricetulus
Reverse Transcription
Saccharomyces cerevisiae
Complementary DNA
peroxisome-targeting signal 1 receptor
Polymerase Chain Reaction
DNA
Genes
Proteins

All Science Journal Classification (ASJC) codes

  • Molecular Biology
  • Cell Biology

これを引用

Peroxisome targeting signal type 1 (PTS1) receptor is involved in import of both PTS1 and PTS2 : Studies with PEX5-defective CHO cell mutants. / Otera, Hidenori; Okumoto, Kanji; Tateishi, Keita; Ikoma, Yuka; Matsuda, Eiko; Nishimura, Maki; Tsukamoto, Toshiro; Osumi, Takashi; Ohashi, Kazumasa; Higuchi, Osamu; Fujiki, Yukio.

:: Molecular and cellular biology, 巻 18, 番号 1, 01.01.1998, p. 388-399.

研究成果: ジャーナルへの寄稿記事

Otera, H, Okumoto, K, Tateishi, K, Ikoma, Y, Matsuda, E, Nishimura, M, Tsukamoto, T, Osumi, T, Ohashi, K, Higuchi, O & Fujiki, Y 1998, 'Peroxisome targeting signal type 1 (PTS1) receptor is involved in import of both PTS1 and PTS2: Studies with PEX5-defective CHO cell mutants', Molecular and cellular biology, 巻. 18, 番号 1, pp. 388-399.
Otera, Hidenori ; Okumoto, Kanji ; Tateishi, Keita ; Ikoma, Yuka ; Matsuda, Eiko ; Nishimura, Maki ; Tsukamoto, Toshiro ; Osumi, Takashi ; Ohashi, Kazumasa ; Higuchi, Osamu ; Fujiki, Yukio. / Peroxisome targeting signal type 1 (PTS1) receptor is involved in import of both PTS1 and PTS2 : Studies with PEX5-defective CHO cell mutants. :: Molecular and cellular biology. 1998 ; 巻 18, 番号 1. pp. 388-399.
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abstract = "To investigate the mechanisms of peroxisome assembly and the molecular basis of peroxisome assembly disorders, we isolated and characterized a peroxisome-deficient CHO cell mutant, ZP139, which was found to belong to human complementation group II, the same group as that of our earlier mutant, ZP105. These mutants had a phenotypic deficiency in the import of peroxisomal targeting signal type 1 (PTS1) proteins. Amino-terminal extension signal (PTS2)-mediated transport, including that of 3-ketoacyl coenzyme A thiolase, was also defective in ZP105 but not in ZP139. PEX5 cDNA, encoding the PTS1 receptor (PTS1R), was isolated from wild-type CHO-K1 cells. PTS1R's deduced primary sequence comprised 595 amino acids, 7 amino acids less than the human homolog, and contained seven tetratricopeptide repeat (TPR) motifs at the C- terminal region. Chinese hamster PTS1R showed 94, 28, and 24{\%} amino acid identity with PTS1Rs from humans, Pichia pastoris, and Saccharomyces cerevisiae, respectively. A PTS1R isoform (PTS1RL) with 632 amino acid residues was identified in CHO cells; for PTSIR, 37 amino acids were inserted between residues at positions 215 and 216 of a shorter isoform (PTS1RS). Southern blot analysis of CHO cell genomic DNA suggested that these two isoforms are derived from a single gene. Both types of PEX5 complemented impaired import of PTS1 in mutants ZP105 and ZP139, PTS2 import in ZP105 was rescued only by PTS1RL. This finding strongly suggests that PTS1RL is also involved in the transport of PTS2. Mutations in PEX5 were determined by reverse transcription-PCR: a G-to-A transition resulted in one amino acid substitution: Gly298Glu of PTS1RS (G335E of PTS1RL) in ZP105 and Gly485Glu of PTS1RS (G522E of PTS1RL) in ZP139. Both mutations were in the TPR domains (TPR1 and TPR6), suggesting the functional consequence of these domains in protein translocation. The implications of these mutations are discussed.",
author = "Hidenori Otera and Kanji Okumoto and Keita Tateishi and Yuka Ikoma and Eiko Matsuda and Maki Nishimura and Toshiro Tsukamoto and Takashi Osumi and Kazumasa Ohashi and Osamu Higuchi and Yukio Fujiki",
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T1 - Peroxisome targeting signal type 1 (PTS1) receptor is involved in import of both PTS1 and PTS2

T2 - Studies with PEX5-defective CHO cell mutants

AU - Otera, Hidenori

AU - Okumoto, Kanji

AU - Tateishi, Keita

AU - Ikoma, Yuka

AU - Matsuda, Eiko

AU - Nishimura, Maki

AU - Tsukamoto, Toshiro

AU - Osumi, Takashi

AU - Ohashi, Kazumasa

AU - Higuchi, Osamu

AU - Fujiki, Yukio

PY - 1998/1/1

Y1 - 1998/1/1

N2 - To investigate the mechanisms of peroxisome assembly and the molecular basis of peroxisome assembly disorders, we isolated and characterized a peroxisome-deficient CHO cell mutant, ZP139, which was found to belong to human complementation group II, the same group as that of our earlier mutant, ZP105. These mutants had a phenotypic deficiency in the import of peroxisomal targeting signal type 1 (PTS1) proteins. Amino-terminal extension signal (PTS2)-mediated transport, including that of 3-ketoacyl coenzyme A thiolase, was also defective in ZP105 but not in ZP139. PEX5 cDNA, encoding the PTS1 receptor (PTS1R), was isolated from wild-type CHO-K1 cells. PTS1R's deduced primary sequence comprised 595 amino acids, 7 amino acids less than the human homolog, and contained seven tetratricopeptide repeat (TPR) motifs at the C- terminal region. Chinese hamster PTS1R showed 94, 28, and 24% amino acid identity with PTS1Rs from humans, Pichia pastoris, and Saccharomyces cerevisiae, respectively. A PTS1R isoform (PTS1RL) with 632 amino acid residues was identified in CHO cells; for PTSIR, 37 amino acids were inserted between residues at positions 215 and 216 of a shorter isoform (PTS1RS). Southern blot analysis of CHO cell genomic DNA suggested that these two isoforms are derived from a single gene. Both types of PEX5 complemented impaired import of PTS1 in mutants ZP105 and ZP139, PTS2 import in ZP105 was rescued only by PTS1RL. This finding strongly suggests that PTS1RL is also involved in the transport of PTS2. Mutations in PEX5 were determined by reverse transcription-PCR: a G-to-A transition resulted in one amino acid substitution: Gly298Glu of PTS1RS (G335E of PTS1RL) in ZP105 and Gly485Glu of PTS1RS (G522E of PTS1RL) in ZP139. Both mutations were in the TPR domains (TPR1 and TPR6), suggesting the functional consequence of these domains in protein translocation. The implications of these mutations are discussed.

AB - To investigate the mechanisms of peroxisome assembly and the molecular basis of peroxisome assembly disorders, we isolated and characterized a peroxisome-deficient CHO cell mutant, ZP139, which was found to belong to human complementation group II, the same group as that of our earlier mutant, ZP105. These mutants had a phenotypic deficiency in the import of peroxisomal targeting signal type 1 (PTS1) proteins. Amino-terminal extension signal (PTS2)-mediated transport, including that of 3-ketoacyl coenzyme A thiolase, was also defective in ZP105 but not in ZP139. PEX5 cDNA, encoding the PTS1 receptor (PTS1R), was isolated from wild-type CHO-K1 cells. PTS1R's deduced primary sequence comprised 595 amino acids, 7 amino acids less than the human homolog, and contained seven tetratricopeptide repeat (TPR) motifs at the C- terminal region. Chinese hamster PTS1R showed 94, 28, and 24% amino acid identity with PTS1Rs from humans, Pichia pastoris, and Saccharomyces cerevisiae, respectively. A PTS1R isoform (PTS1RL) with 632 amino acid residues was identified in CHO cells; for PTSIR, 37 amino acids were inserted between residues at positions 215 and 216 of a shorter isoform (PTS1RS). Southern blot analysis of CHO cell genomic DNA suggested that these two isoforms are derived from a single gene. Both types of PEX5 complemented impaired import of PTS1 in mutants ZP105 and ZP139, PTS2 import in ZP105 was rescued only by PTS1RL. This finding strongly suggests that PTS1RL is also involved in the transport of PTS2. Mutations in PEX5 were determined by reverse transcription-PCR: a G-to-A transition resulted in one amino acid substitution: Gly298Glu of PTS1RS (G335E of PTS1RL) in ZP105 and Gly485Glu of PTS1RS (G522E of PTS1RL) in ZP139. Both mutations were in the TPR domains (TPR1 and TPR6), suggesting the functional consequence of these domains in protein translocation. The implications of these mutations are discussed.

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