Peroxisome-deficient Chinese hamster ovary cells with point mutations in peroxisome assembly factor-1.
Chinese hamster ovary (CHO) mutant cells deficient in peroxisome biogenesis regain peroxisomes after transfection with a cDNA coding for peroxisome assembly factor (PAF)-1 from rat liver. Reconstitution of the transfected mutant cells with wild-type cytoplasm was not required, demonstrating that expression of the PAF-1 gene alone was sufficient for the restoration of peroxisome biogenesis. Plasmalogen biosynthesis in the transfected mutants was also restored to approximately wild-type levels. The nucleotide sequence of the cDNA encoding the open reading frame for PAF-1 from CHO-K1 cells was determined. This allowed us to identify point mutations of PAF-1 in two peroxisomal mutant cell lines. The mutation in ZR-78 cells changed a cysteine to a tyrosine codon in a region located at the carboxyl terminus of the protein, which resembles the zinc finger motif of DNA-binding proteins. A point mutation in the PAF-1 gene of ZR-82 leads to premature termination.
Thieringer
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Merck Research Laboratories, Rahway, New Jersey 07065.
Raetz
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CR
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D016466:CHO Cells; D002374:Catalase / Q000032:analysis / Q000378:metabolism; D002462:Cell Membrane / Q000378:metabolism; D003001:Cloning, Molecular; D003062:Codon / Q000235:genetics; D006224:Cricetinae; D004247:DNA; D004252:DNA Mutational Analysis; D006801:Humans; D008099:Liver / Q000378:metabolism; D008565:Membrane Proteins / Q000096:biosynthesis* / Q000235:genetics; D008830:Microbodies / Q000378:metabolism*; D008969:Molecular Sequence Data; D016296:Mutagenesis; D000074428:Peroxisomal Biogenesis Factor 2; D017354:Point Mutation; D011061:Poly A / Q000302:isolation & purification / Q000378:metabolism; D012313:RNA / Q000302:isolation & purification / Q000378:metabolism; D012333:RNA, Messenger / Q000302:isolation & purification / Q000378:metabolism; D051381:Rats; D017386:Sequence Homology, Amino Acid; D014443:Tyrosine
D003062:Codon; D008565:Membrane Proteins; D012333:RNA, Messenger; D000074428:Peroxisomal Biogenesis Factor 2; D011061:Poly A; D014443:Tyrosine; D012313:RNA; D004247:DNA; D002374:Catalase