R. Toyama et al., Isolation, characterization and mutation analysis of PEX13-defective Chinese hamster ovary cell mutants, HUM MOL GEN, 8(9), 1999, pp. 1673-1681
We isolated peroxisome biogenesis mutants ZP128 and ZP150 from rat PEX2-tra
nsformed Chinese hamster ovary (CHO) cells, by the 9-(1'-pyrene)nonanol/ult
raviolet method. The mutants lacked morphologically recognizable peroxisome
s and showed a typical peroxisome assembly-defective phenotype such as a hi
gh sensitivity to 12-(1'-pyrene)dodecanoic acid/UV treatment. By means of P
EX 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 i
solated; its deduced sequence comprises 405 amino acids with 93% identity t
o 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 o
ne 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 phe
notype of impaired PEX13.