Isolation and characterization of peroxisome-deficient Chinese hamster ovary cell mutants representing human complementation group III

Kanji Okumoto, Akemi Bogaki, Keita Tateishi, Toshiro Tsukamoto, Takashi Osumi, Nobuyuki Shimozawa, Yasuyuki Suzuki, Tadao Orii, Yukio Fujiki

研究成果: ジャーナルへの寄稿記事

38 引用 (Scopus)

抄録

We made use of the 9-(1'-pyrene)nonanol/ultraviolet (P9OH/UV) method and isolated peroxisome-deficient mutant cells. TKa cells, the wild-type Chinese hamster ovary (CHO) cells, CHO-K1, that had been stably transfected with cDNA encoding Pex2p (formerly peroxisome assembly factor-1, PAF-1) were used to avoid frequent isolation of the Z65-type, PEX2-defective mutants. P9OH/UV- resistant cell colonies were examined for the intracellular location of catalase, a peroxisomal matrix enzyme, by immunofluorescence microscopy and using anti-catalase antibody. As six mutant cell clones showed cytosolic catalase, there was likely to be a deficiency in peroxisome assembly. These mutants also showed the typical peroxisome assembly-defective phenotype, including significant decrease of dihydroxyacetonephosphate acyltransferase, the first step key enzyme in plasmalogen synthesis, and loss of resistance to 12-(1'-pyrene)dodecanoic acid/UV treatment. By transfection of Pex2p and Pex6p (formerly PAF-2) cDNAs and cell fusion analysis between the CHO cell mutants, two mutants, ZP104 and ZP109, were found to belong to a novel complementation group. Further complementation analysis using fibroblasts from patients with peroxisome biogenesis disorders revealed that the mutants belonged to human complementation group III. Taken together, ZP104 and ZP109 are in a newly identified fifth complementation group in CHO mutants reported to date and represent the human complementation group III.

元の言語英語
ページ(範囲)11-20
ページ数10
ジャーナルExperimental Cell Research
233
発行部数1
DOI
出版物ステータス出版済み - 5 25 1997

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Peroxisomes
Cricetulus
Ovary
Catalase
glycerone-phosphate O-acyltransferase
Complementary DNA
Plasmalogens
Cell Fusion
Enzymes
Fluorescence Microscopy
Transfection
Anti-Idiotypic Antibodies
Clone Cells
Fibroblasts
Phenotype
9-(1'-pyrene)nonanol

All Science Journal Classification (ASJC) codes

  • Cell Biology

これを引用

Isolation and characterization of peroxisome-deficient Chinese hamster ovary cell mutants representing human complementation group III. / Okumoto, Kanji; Bogaki, Akemi; Tateishi, Keita; Tsukamoto, Toshiro; Osumi, Takashi; Shimozawa, Nobuyuki; Suzuki, Yasuyuki; Orii, Tadao; Fujiki, Yukio.

:: Experimental Cell Research, 巻 233, 番号 1, 25.05.1997, p. 11-20.

研究成果: ジャーナルへの寄稿記事

Okumoto, Kanji ; Bogaki, Akemi ; Tateishi, Keita ; Tsukamoto, Toshiro ; Osumi, Takashi ; Shimozawa, Nobuyuki ; Suzuki, Yasuyuki ; Orii, Tadao ; Fujiki, Yukio. / Isolation and characterization of peroxisome-deficient Chinese hamster ovary cell mutants representing human complementation group III. :: Experimental Cell Research. 1997 ; 巻 233, 番号 1. pp. 11-20.
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abstract = "We made use of the 9-(1'-pyrene)nonanol/ultraviolet (P9OH/UV) method and isolated peroxisome-deficient mutant cells. TKa cells, the wild-type Chinese hamster ovary (CHO) cells, CHO-K1, that had been stably transfected with cDNA encoding Pex2p (formerly peroxisome assembly factor-1, PAF-1) were used to avoid frequent isolation of the Z65-type, PEX2-defective mutants. P9OH/UV- resistant cell colonies were examined for the intracellular location of catalase, a peroxisomal matrix enzyme, by immunofluorescence microscopy and using anti-catalase antibody. As six mutant cell clones showed cytosolic catalase, there was likely to be a deficiency in peroxisome assembly. These mutants also showed the typical peroxisome assembly-defective phenotype, including significant decrease of dihydroxyacetonephosphate acyltransferase, the first step key enzyme in plasmalogen synthesis, and loss of resistance to 12-(1'-pyrene)dodecanoic acid/UV treatment. By transfection of Pex2p and Pex6p (formerly PAF-2) cDNAs and cell fusion analysis between the CHO cell mutants, two mutants, ZP104 and ZP109, were found to belong to a novel complementation group. Further complementation analysis using fibroblasts from patients with peroxisome biogenesis disorders revealed that the mutants belonged to human complementation group III. Taken together, ZP104 and ZP109 are in a newly identified fifth complementation group in CHO mutants reported to date and represent the human complementation group III.",
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AU - Osumi, Takashi

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AU - Suzuki, Yasuyuki

AU - Orii, Tadao

AU - Fujiki, Yukio

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