Kinetic analysis of genetic complementation in heterokaryons of propionyl CoA carboxylase-deficient human fibroblasts

B. Wolf, H. F. Willard, L. E. Rosenberg

Research output: Contribution to journalArticle

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Abstract

The authors studied genetic complementation of propionyl CoA carboxylase (PCC) deficiency in cultures of polyethylene glycol (PEG)-induced heterokaryons, using mutant fibroblast lines assigned to 5 mutant classes, designated bio, pcc A, pcc B, pcc C, and pcc BC. By measuring PCC activity directly in extracts of fused cells or indirectly in intact cells by [1-14C]propionate utilization, the authors confirmed the nonlinear nature of the PCC deficiency complementation map described by Gravel et al. When they studied the kinetics of complementation, they detected 3 distinct patterns using the [1-14C]propionate utilization assay. When either pcc A or pcc C lines were fused to bio cells, 14C-fixation increased to half of the maximally restored values within 4 hr. In pcc A x pcc C crosses or in pcc A x pcc B crosses, however, complementation was much slower. In fusions between pcc B and pcc C cells, a third pattern was elicited; complementation was incomplete, maximum restoration of PCC activity being <20% of that observed in other complementing crosses. From these data and previous biochemical evidence, the authors suggest: (1) that the bio and pcc mutations affect different genes; (2) that complementation between pcc A and either pcc B, pcc C, or pcc BC lines is intergenic and involves subunit exchange and synthesis of new PCC molecules; and (3) that complementation between pcc B and pcc C mutants is interallelic.

Original languageEnglish (US)
Pages (from-to)16-25
Number of pages10
JournalAmerican Journal of Human Genetics
Volume32
Issue number1
StatePublished - Jan 1 1980

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Methylmalonyl-CoA Decarboxylase
Propionic Acidemia
Fibroblasts
Propionates
Cell Extracts
Mutation
Genes

ASJC Scopus subject areas

  • Genetics

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title = "Kinetic analysis of genetic complementation in heterokaryons of propionyl CoA carboxylase-deficient human fibroblasts",
abstract = "The authors studied genetic complementation of propionyl CoA carboxylase (PCC) deficiency in cultures of polyethylene glycol (PEG)-induced heterokaryons, using mutant fibroblast lines assigned to 5 mutant classes, designated bio, pcc A, pcc B, pcc C, and pcc BC. By measuring PCC activity directly in extracts of fused cells or indirectly in intact cells by [1-14C]propionate utilization, the authors confirmed the nonlinear nature of the PCC deficiency complementation map described by Gravel et al. When they studied the kinetics of complementation, they detected 3 distinct patterns using the [1-14C]propionate utilization assay. When either pcc A or pcc C lines were fused to bio cells, 14C-fixation increased to half of the maximally restored values within 4 hr. In pcc A x pcc C crosses or in pcc A x pcc B crosses, however, complementation was much slower. In fusions between pcc B and pcc C cells, a third pattern was elicited; complementation was incomplete, maximum restoration of PCC activity being <20{\%} of that observed in other complementing crosses. From these data and previous biochemical evidence, the authors suggest: (1) that the bio and pcc mutations affect different genes; (2) that complementation between pcc A and either pcc B, pcc C, or pcc BC lines is intergenic and involves subunit exchange and synthesis of new PCC molecules; and (3) that complementation between pcc B and pcc C mutants is interallelic.",
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Kinetic analysis of genetic complementation in heterokaryons of propionyl CoA carboxylase-deficient human fibroblasts. / Wolf, B.; Willard, H. F.; Rosenberg, L. E.

In: American Journal of Human Genetics, Vol. 32, No. 1, 01.01.1980, p. 16-25.

Research output: Contribution to journalArticle

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