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. 1977 Jul;29(4):378–388.

Genetic complementation of propionyl-CoA carboxylase deficiency in cultured human fibroblasts.

R A Gravel, K F Lam, K J Scully, Y Hsia
PMCID: PMC1685391  PMID: 195466

Abstract

Propionyl-CoA carboxylase (PCC) deficiency is an inherited metabolic disorder showing considerable variability of expression. We have investigated the possibility that there is a genetic basis for the clinical heterogeneity in this disorder by examining complementation in Sendai virus mediated heterokaryons of mutant fibroblast strains. Restoration of PCC activity was monitored in individual multinucleate cells in situ using a radioautographic procedure which detects the incorporation of 14C-propionate into trichloracetic acid precipitable material. Each mutant strain incorporated negligible amounts of radioactivity compared to control strains. Activity was not restored when different mutants were mixed without virus or when homokaryons were produced by self-fusion. Seven mutant strains were fused in all pairwise combinations and examined for increased 14C-propionate incorporation in heterokaryons. Two main complementation groups were revealed. One group was composed of three mutants. The other was a complex group composed of four mutants in which intragroup complementation was demonstrated. Two mutants showing excellent complementation by radioautography were examined for complementation by the direct assay of PCC ACTIVITY. The enzyme activity of virus-treated preparations with 23% multinucleate cells was 183 U (pmol/min/mg protein) compared to 16 U for the untreated mixture (normal range 450-850 u). We conclude that PCC deficiency resulted from mutations of heterogeneous origin, although the classification of mutants into complementation groups did not correlate with patterns of clinical heterogeneity.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Ando T., Rasmussen K., Nyhan W. L., Donnell G. N., Barnes N. D. Propionic acidemia in patients with ketotic hyperglycinemia. J Pediatr. 1971 May;78(5):827–832. doi: 10.1016/s0022-3476(71)80354-2. [DOI] [PubMed] [Google Scholar]
  2. Barnes N. D., Hull D., Balgobin L., Gompertz D. Biotin-responsive propionicacidaemia. Lancet. 1970 Aug 1;2(7666):244–245. doi: 10.1016/s0140-6736(70)92590-0. [DOI] [PubMed] [Google Scholar]
  3. Bartlett K., Gompertz D. Combined carboxylase defect: biotin-responsiveness in cultured fibroblasts. Lancet. 1976 Oct 9;2(7989):804–804. doi: 10.1016/s0140-6736(76)90640-1. [DOI] [PubMed] [Google Scholar]
  4. Brandt I. K., Hsia Y. E., Clement D. H., Provence S. A. Propionicacidemia (ketotic hyperglycinemia): dietary treatment resulting in normal growth and development. Pediatrics. 1974 Mar;53(3):391–395. [PubMed] [Google Scholar]
  5. Callahan J. W., Khalil M., Philippart M. Sphingomyelinases in human tissues. II. Absence of a specific enzyme from liver and brain of Niemann-Pick disease, type C. Pediatr Res. 1975 Dec;9(12):908–913. doi: 10.1203/00006450-197512000-00009. [DOI] [PubMed] [Google Scholar]
  6. Callahan J. W., Khalil M. Sphingomyelinases in human tissues. III. Expression of Niemann-Pick disease in cultured skin fibroblasts. Pediatr Res. 1975 Dec;9(12):914–918. doi: 10.1203/00006450-197512000-00010. [DOI] [PubMed] [Google Scholar]
  7. DULBECCO R., VOGT M. Plaque formation and isolation of pure lines with poliomyelitis viruses. J Exp Med. 1954 Feb;99(2):167–182. doi: 10.1084/jem.99.2.167. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Giorgio A. J., Whitaker T. R. Some properties of propionyl CoA carboxylase partially purified from human liver. Biochem Med. 1973 Jun;7(3):473–478. doi: 10.1016/0006-2944(73)90069-0. [DOI] [PubMed] [Google Scholar]
  9. Gompertz D., Goodey P. A., Thom H., Russell G., Johnston A. W., Mellor D. H., MacLean M. W., Ferguson-Smith M. E., Ferguson-Smith M. A. Prenatal diagnosis and family studies in a case of propionicacidaemia. Clin Genet. 1975 Oct;8(4):244–250. doi: 10.1111/j.1399-0004.1975.tb01499.x. [DOI] [PubMed] [Google Scholar]
  10. Gompertz D., Storrs C. N., Bau D. C., Peters T. J., Hughes E. A. Localisation of enzymic defect in propionicacidaemia. Lancet. 1970 May 30;1(7657):1140–1143. doi: 10.1016/s0140-6736(70)91216-x. [DOI] [PubMed] [Google Scholar]
  11. Gravel R. A., Mahoney M. J., Ruddle F. H., Rosenberg L. E. Genetic complementation in heterokaryons of human fibroblasts defective in cobalamin metabolism. Proc Natl Acad Sci U S A. 1975 Aug;72(8):3181–3185. doi: 10.1073/pnas.72.8.3181. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Guibaud P., Divry P., Marcon G., Cotton J. B., Collombel C., Larbre F. Une forme a révélation tardive de l'acidemie propionique. Arch Fr Pediatr. 1975 Mar;32(3):259–273. [PubMed] [Google Scholar]
  13. Hill H. Z., Goodman S. I. Detection of inborn errors of metabolism. III. Defects in urea cycle metabolism. Clin Genet. 1974;6(2):79–81. [PubMed] [Google Scholar]
  14. Hsia Y. E., Scully K. J., Rosenberg L. E. Defective propionate carboxylation in ketotic hyperglycinaemia. Lancet. 1969 Apr 12;1(7598):757–758. doi: 10.1016/s0140-6736(69)91757-7. [DOI] [PubMed] [Google Scholar]
  15. Hsia Y. E., Scully K. J., Rosenberg L. E. Inherited propionyl-Coa carboxylase deficiency in "ketotic hyperglycinemia". J Clin Invest. 1971 Jan;50(1):127–130. doi: 10.1172/JCI106466. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Kraemer K. H., De Weerd-Kastelein E. A., Robbins J. H., Keijzer W., Barrett S. F., Petinga R. A., Bootsma D. Five complementation groups in xeroderma pigmentosum. Mutat Res. 1975 Dec;33(2-3):327–340. doi: 10.1016/0027-5107(75)90208-0. [DOI] [PubMed] [Google Scholar]
  17. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  18. Lyons L. B., Cox R. P., Dancis J. Complementation analysis of maple syrup urine disease in heterokaryons derived from cultured human fibroblasts. Nature. 1973 Jun 29;243(5409):533–535. doi: 10.1038/243533a0. [DOI] [PubMed] [Google Scholar]
  19. Rattazzi M. C., Brown J. A., Davidson R. G., Shows T. B. Studies on complementation of beta hexosaminidase deficiency in human GM2 gangliosidosis. Am J Hum Genet. 1976 Mar;28(2):143–154. [PMC free article] [PubMed] [Google Scholar]
  20. Seegmiller J. E. Inherited deficiency of hypoxanthine-guanine phosphoribosyltransferase in X-linked uric aciduria (the Lesch-Nyhan syndrome and its variants). Adv Hum Genet. 1976;6:75–163. doi: 10.1007/978-1-4615-8264-9_2. [DOI] [PubMed] [Google Scholar]
  21. Stanners C. P., Eliceiri G. L., Green H. Two types of ribosome in mouse-hamster hybrid cells. Nat New Biol. 1971 Mar 10;230(10):52–54. doi: 10.1038/newbio230052a0. [DOI] [PubMed] [Google Scholar]
  22. Wadlington W. B., Kilroy A., Ando T., Sweetman L., Nyhan W. L. Hyperglycinemia and propionyl coA carboxylase deficiency and episodic severe illness without consistent ketosis. J Pediatr. 1975 May;86(5):707–712. doi: 10.1016/s0022-3476(75)80354-4. [DOI] [PubMed] [Google Scholar]

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