Complementation Studies of Isovaleric Acidemia and Glutaric Aciduria Type II Using Cultured Skin Fibroblasts

Overview

Journal J Clin Invest

Specialty General Medicine

Date 1983 Nov 1

PMID 6630517

Citations 5

Authors

B Dubiel

C Dabrowski

R Wetts

K Tanaka

Affiliations

Soon will be listed here.

Abstract

Using cultured skin fibroblasts, we studied the heterogeneity of inborn errors of leucine metabolism such as isovaleric acidemia (IVA), glutaric aciduria type II (GA II), and multiple carboxylase deficiency (MC). We first developed a simple macromolecular-labeling test to measure the ability of cells to oxidize [1-14C]isovaleric acid in situ in culture. Cells from two different lines were fused using polyethylene glycol, and the ability of the heterokaryons to oxidize [1-14C]isovaleric acid was tested by the macromolecular-labeling test. The MC line complemented with all 10 IVA lines tested; heterokaryons showed 99 +/- 68% more activity than the unfused mixture of component cells. GA II/IVA heterokaryons exhibited poor growth, but when the culture remained confluent, the GA II cells complemented with all six IVA lines tested, showing a 71 +/- 41% increase in activity. The relatively large standard deviations are due to a few experiments in which significant enhancement of macromolecular-labeling test activity was not observed upon fusion, but significant complementation was clearly observed in repeats of the same combinations. These results are consistent with our previous findings, which indicated that the decreased ability of GA II cells to oxidize isovaleryl-CoA involves a defective electron-transporting system rather than a defective isovaleryl-CoA dehydrogenase. IVA/IVA heterokaryons showed no complementation in any combination tested, indicating no detectable heterogeneity in isovaleric acidemia. This finding indicates that the same gene is mutated in all IVA lines. Previous results indicated that this gene codes for isovaleryl-CoA dehydrogenase.

Citing Articles

Genetic complementation analysis of 3-hydroxy-3-methylglutaryl-coenzyme A lyase deficiency in cultured fibroblasts.

Sovik O, Sweetman L, Gibson K, Nyhan W Am J Hum Genet. 1984; 36(4):791-801.

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L-carnitine and glycine therapy in isovaleric acidaemia.

Chalmers R, De Sousa C, Tracey B, Stacey T, Weaver C, Bradley D J Inherit Metab Dis. 1985; 8 Suppl 2:141-2.

PMID: 3930869 DOI: 10.1007/BF01811499.

Molecular heterogeneity of variant isovaleryl-CoA dehydrogenase from cultured isovaleric acidemia fibroblasts.

Ikeda Y, Keese S, Tanaka K Proc Natl Acad Sci U S A. 1985; 82(20):7081-5.

PMID: 3863140 PMC: 391314. DOI: 10.1073/pnas.82.20.7081.

Prenatal diagnosis of 3-hydroxy-3-methylglutaric aciduria by GC-MS and enzymology on cultured amniocytes and chorionic villi.

Chalmers R, Tracey B, Mistry J, Stacey T, McFadyen I J Inherit Metab Dis. 1989; 12(3):286-92.

PMID: 2482386 DOI: 10.1007/BF01799219.

Nucleotide sequence of messenger RNA encoding human isovaleryl-coenzyme A dehydrogenase and its expression in isovaleric acidemia fibroblasts.

Matsubara Y, Ito M, Glassberg R, Satyabhama S, Ikeda Y, Tanaka K J Clin Invest. 1990; 85(4):1058-64.

PMID: 2318964 PMC: 296535. DOI: 10.1172/JCI114536.

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