PEX12, the pathogenic gene of group III Zellweger syndrome: cDNA cloning by functional complementation on a CHO cell mutant, patient analysis, and characterization of Pex12p

Kanji Okumoto, Nobuyuki Shimozawa, Atsusi Kawai, Shigehiko Tamura, Toshiro Tsukamoto, Takashi Osumi, Hugo Moser, Ronald J.A. Wanders, Yasuyuki Suzuki, Naomi Kondo, Yukio Fujiki

研究成果: ジャーナルへの寄稿記事

89 引用 (Scopus)

抄録

Rat PEX12 cDNA was isolated by functional complementation of peroxisome deficiency of a mutant CHO cell line, ZP109 (K. Okumoto, A. Bogaki, K. Tatelshi, T. Tsukamoto, T. Osumi, N. Shimozawa, Y. Suzuki, T. Orii, and Y. Fujiki, Exp. Cell Res. 233:11-20, 1997), using a transient transfection assay and an ectopic, readily visible marker, green fluorescent protein. This cDNA encodes a 359-amino-acid membrane protein of peroxisomes with two transmembrane segment and a cysteine-rich zinc finger, the RING motif. A stable transformant of ZP109 with the PEX12 was morphologically and biochemically restored for peroxisome biogenesis. Pex12p was shown by expression of bona fide as well as epitope-tagged Pex12p to expose both N- and C-terminal regions to the cytosol. Fibroblasts derived from patients with the perosizome deficiency Zellweger syndrome of complementation group III (CG-III) were also complemented for peroxisome biogenesis with PEX12. Two unrelated patients of this group manifesting peroxisome deficiency disorders possessed homozygous, inactivating PEX12 mutations: in one, Arg180Thr by one point mutation, and in the other, deletion of two nucleotides in codons for291 Asn and 292Ser, creating an apparently unchanged codon for Asn and a codon 292 for termination. These results indicate that the gene encoding peroxisome assembly factor Pex12p is a pathogenic gene of CG-III peroxisome deficiency. Moreover, truncation and site mutation studies, including patient PEX12 analysis, demonstrated that the cytoplasmically oriented N- and C-terminal parts of Pex12p are essential for biological function.

元の言語英語
ページ(範囲)4324-4336
ページ数13
ジャーナルMolecular and cellular biology
18
発行部数7
DOI
出版物ステータス出版済み - 7 1998

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Zellweger Syndrome
Peroxisomes
CHO Cells
Organism Cloning
Complementary DNA
Genes
Codon
Mutation
Terminator Codon
Zinc Fingers
Green Fluorescent Proteins
Point Mutation
Cytosol
Transfection
Cysteine
Epitopes
Membrane Proteins
Nucleotides
Fibroblasts
Amino Acids

All Science Journal Classification (ASJC) codes

  • Molecular Biology
  • Cell Biology

これを引用

PEX12, the pathogenic gene of group III Zellweger syndrome : cDNA cloning by functional complementation on a CHO cell mutant, patient analysis, and characterization of Pex12p. / Okumoto, Kanji; Shimozawa, Nobuyuki; Kawai, Atsusi; Tamura, Shigehiko; Tsukamoto, Toshiro; Osumi, Takashi; Moser, Hugo; Wanders, Ronald J.A.; Suzuki, Yasuyuki; Kondo, Naomi; Fujiki, Yukio.

:: Molecular and cellular biology, 巻 18, 番号 7, 07.1998, p. 4324-4336.

研究成果: ジャーナルへの寄稿記事

Okumoto, Kanji ; Shimozawa, Nobuyuki ; Kawai, Atsusi ; Tamura, Shigehiko ; Tsukamoto, Toshiro ; Osumi, Takashi ; Moser, Hugo ; Wanders, Ronald J.A. ; Suzuki, Yasuyuki ; Kondo, Naomi ; Fujiki, Yukio. / PEX12, the pathogenic gene of group III Zellweger syndrome : cDNA cloning by functional complementation on a CHO cell mutant, patient analysis, and characterization of Pex12p. :: Molecular and cellular biology. 1998 ; 巻 18, 番号 7. pp. 4324-4336.
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title = "PEX12, the pathogenic gene of group III Zellweger syndrome: cDNA cloning by functional complementation on a CHO cell mutant, patient analysis, and characterization of Pex12p",
abstract = "Rat PEX12 cDNA was isolated by functional complementation of peroxisome deficiency of a mutant CHO cell line, ZP109 (K. Okumoto, A. Bogaki, K. Tatelshi, T. Tsukamoto, T. Osumi, N. Shimozawa, Y. Suzuki, T. Orii, and Y. Fujiki, Exp. Cell Res. 233:11-20, 1997), using a transient transfection assay and an ectopic, readily visible marker, green fluorescent protein. This cDNA encodes a 359-amino-acid membrane protein of peroxisomes with two transmembrane segment and a cysteine-rich zinc finger, the RING motif. A stable transformant of ZP109 with the PEX12 was morphologically and biochemically restored for peroxisome biogenesis. Pex12p was shown by expression of bona fide as well as epitope-tagged Pex12p to expose both N- and C-terminal regions to the cytosol. Fibroblasts derived from patients with the perosizome deficiency Zellweger syndrome of complementation group III (CG-III) were also complemented for peroxisome biogenesis with PEX12. Two unrelated patients of this group manifesting peroxisome deficiency disorders possessed homozygous, inactivating PEX12 mutations: in one, Arg180Thr by one point mutation, and in the other, deletion of two nucleotides in codons for291 Asn and 292Ser, creating an apparently unchanged codon for Asn and a codon 292 for termination. These results indicate that the gene encoding peroxisome assembly factor Pex12p is a pathogenic gene of CG-III peroxisome deficiency. Moreover, truncation and site mutation studies, including patient PEX12 analysis, demonstrated that the cytoplasmically oriented N- and C-terminal parts of Pex12p are essential for biological function.",
author = "Kanji Okumoto and Nobuyuki Shimozawa and Atsusi Kawai and Shigehiko Tamura and Toshiro Tsukamoto and Takashi Osumi and Hugo Moser and Wanders, {Ronald J.A.} and Yasuyuki Suzuki and Naomi Kondo and Yukio Fujiki",
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T1 - PEX12, the pathogenic gene of group III Zellweger syndrome

T2 - cDNA cloning by functional complementation on a CHO cell mutant, patient analysis, and characterization of Pex12p

AU - Okumoto, Kanji

AU - Shimozawa, Nobuyuki

AU - Kawai, Atsusi

AU - Tamura, Shigehiko

AU - Tsukamoto, Toshiro

AU - Osumi, Takashi

AU - Moser, Hugo

AU - Wanders, Ronald J.A.

AU - Suzuki, Yasuyuki

AU - Kondo, Naomi

AU - Fujiki, Yukio

PY - 1998/7

Y1 - 1998/7

N2 - Rat PEX12 cDNA was isolated by functional complementation of peroxisome deficiency of a mutant CHO cell line, ZP109 (K. Okumoto, A. Bogaki, K. Tatelshi, T. Tsukamoto, T. Osumi, N. Shimozawa, Y. Suzuki, T. Orii, and Y. Fujiki, Exp. Cell Res. 233:11-20, 1997), using a transient transfection assay and an ectopic, readily visible marker, green fluorescent protein. This cDNA encodes a 359-amino-acid membrane protein of peroxisomes with two transmembrane segment and a cysteine-rich zinc finger, the RING motif. A stable transformant of ZP109 with the PEX12 was morphologically and biochemically restored for peroxisome biogenesis. Pex12p was shown by expression of bona fide as well as epitope-tagged Pex12p to expose both N- and C-terminal regions to the cytosol. Fibroblasts derived from patients with the perosizome deficiency Zellweger syndrome of complementation group III (CG-III) were also complemented for peroxisome biogenesis with PEX12. Two unrelated patients of this group manifesting peroxisome deficiency disorders possessed homozygous, inactivating PEX12 mutations: in one, Arg180Thr by one point mutation, and in the other, deletion of two nucleotides in codons for291 Asn and 292Ser, creating an apparently unchanged codon for Asn and a codon 292 for termination. These results indicate that the gene encoding peroxisome assembly factor Pex12p is a pathogenic gene of CG-III peroxisome deficiency. Moreover, truncation and site mutation studies, including patient PEX12 analysis, demonstrated that the cytoplasmically oriented N- and C-terminal parts of Pex12p are essential for biological function.

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