TY - JOUR
T1 - Isolation, characterization and mutation analysis of PEX13-defective Chinese hamster ovary cell mutants
AU - Toyama, Ryusuke
AU - Mukai, Satoru
AU - Itagaki, Atsushi
AU - Tamura, Shigehiko
AU - Shimozawa, Nobuyuki
AU - Suzuki, Yasuyuki
AU - Kondo, Naomi
AU - Wanders, Ronald J.A.
AU - Fujiki, Yukio
N1 - Funding Information:
We thank K. Tateishi for participating in the initial stage of this work, A. Tamura for technical assistance, K. Okumoto, M. Honsho, T. Tsukamoto and T. Osumi for comments, and R. Tanaka for secretarial work. This work was supported in part by a CREST grant (to Y.F.) from the Japan Science and Technology Corporation; Grants-in-Aid for Scientific Research (Y.F.) from the Ministry of Education, Science, Sports and Culture of Japan.
PY - 1999
Y1 - 1999
N2 - We isolated peroxisome biogenesis mutants ZP128 and ZP150 from rat PEX2-transformed Chinese hamster ovary (CHO) cells, by the 9-(1'-pyrene)nonanol/ultraviolet method. The mutants lacked morphologically recognizable peroxisomes and showed a typical peroxisome assembly-defective phenotype such as a high sensitivity to 12-(1'-pyrene)dodecanoic acid/UV treatment. By means of PEX cDNA transfection and cell fusion, ZP128 and ZP150 were found to belong to a recently identified complementation group H. Expression of human PEX13 cDNA restored peroxisome assembly in ZP128 and ZP150. CHO cell PEX13 was isolated; its deduced sequence comprises 405 amino acids with 93% identity to human Pex13p. Mutation in PEX13 of mutant ZP150 was determined by RT-PCR: G to A transition resulted in one amino acid substitution, Ser319Asn, in one allele and truncation of a 42 amino acid sequence from Asp265 to Lys306 in another allele. Therefore, ZP128 and ZP150 are CHO cell lines with a phenotype of impaired PEX13.
AB - We isolated peroxisome biogenesis mutants ZP128 and ZP150 from rat PEX2-transformed Chinese hamster ovary (CHO) cells, by the 9-(1'-pyrene)nonanol/ultraviolet method. The mutants lacked morphologically recognizable peroxisomes and showed a typical peroxisome assembly-defective phenotype such as a high sensitivity to 12-(1'-pyrene)dodecanoic acid/UV treatment. By means of PEX cDNA transfection and cell fusion, ZP128 and ZP150 were found to belong to a recently identified complementation group H. Expression of human PEX13 cDNA restored peroxisome assembly in ZP128 and ZP150. CHO cell PEX13 was isolated; its deduced sequence comprises 405 amino acids with 93% identity to human Pex13p. Mutation in PEX13 of mutant ZP150 was determined by RT-PCR: G to A transition resulted in one amino acid substitution, Ser319Asn, in one allele and truncation of a 42 amino acid sequence from Asp265 to Lys306 in another allele. Therefore, ZP128 and ZP150 are CHO cell lines with a phenotype of impaired PEX13.
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U2 - 10.1093/hmg/8.9.1673
DO - 10.1093/hmg/8.9.1673
M3 - Article
C2 - 10441330
AN - SCOPUS:0032832999
SN - 0964-6906
VL - 8
SP - 1673
EP - 1681
JO - Human Molecular Genetics
JF - Human Molecular Genetics
IS - 9
ER -