ANALYSIS OF MUTATIONS IN ALLELES OF THE FUR GENE FROM AN ENDOPROTEASE-DEFICIENT CHINESE-HAMSTER OVARY CELL STRAIN

被引:37
作者
SPENCE, MJ [1 ]
SUCIC, JF [1 ]
FOLEY, BT [1 ]
MOEHRING, TJ [1 ]
机构
[1] UNIV VERMONT,VERMONT CANC CTR,BURLINGTON,VT 05405
关键词
D O I
10.1007/BF02255818
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
RPE.40 mutant cells differ from wild-type Chinese hamster ovary (CHO-K1) cells in their increased resistance to Pseudomonas exotoxin A and their inability to process the insulin proreceptor and certain viral envelope proproteins. Northern analysis revealed that RPE.40 cells maintained a substantially lower steady-state level of 4.0 kb fur mRNA than did CHO-K1 cells. Analysis of fur cDNAs showed that RPE.40 cells were diploid at the fur locus, and RPE.40 cells had a Cys (TGC) to Tyr (TAG) mutation in codon 196 of one allele (allele I). Approximately 25-30% ofthe CHO-K1 cells wer e also heterozygous (Tyr/Cys) at codon 196, and pre-mRNAs transcribed from the second allele (allele II) in RPE.40 cells were defectively spliced. All other pre-mRNAs were correctly spliced. Rapid turnover of defectively spliced transcripts may account for the reduced steady-state level of fur mRNA observed in RPE.40 cells. Our results provide a mechanistic basis for the endoprotease-deficient phenotype of RPE.40 cells.
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页码:1 / 18
页数:18
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