Gene conversion in salt-losing congenital adrenal hyperplasia with absent complement C4B protein

P. A. Donohoue, C. van Dop, R. H. McLean, P. C. White, N. Jospe, C. J. Migeon

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Abstract

Two of four siblings expressed the salt-losing form of congenital adrenal hyperplasia due to 21-hydroxylase deficiency (CAH) and had identical human lymphocyte antigen (HLA) and complement C4 (fourth component of complement) types (HLA-A3, C4, B35, C4A3, C4BQO, DR1/A2, C-,B18, C4A3, C4BQO,DR6). The father and one unaffected sibling were heterozygous carriers of CAH, as determined by a 30-min iv ACTH stimulation test and HLA typing. In addition, the iv ACTH stimulation test revealed that the mother and the other unaffected sibling also carried an allele for an attenuated form of CAH. Restriction endonuclease digests of genomic DNA obtained from members of this family and from normal unrelated subjects were hybridized with cDNA probes encoding human 21-hydroxylase and C4. With the 21-hydroxylase probe, Southern blots prepared from control DNA samples revealed two major restriction fragments in each of four restriction endonuclease digests; TaqI produced major bands at 3.7 and 3.2 kilobases (kb), KpnI at 4.0 and 2.9 kb, EcoRI at 18 and 13 kb, and BglII at 15 and 12.5 kb. Southern blots prepared from DNA of the two patients lacked the 3.7-kb TaqI and 2.9-kb KpnI fragments, but had increased hybridization intensity (relative to control DNA samples) in the 3.2-kb TaqI and 4.0-kb KpnI fragments. By contrast, blots with EcoRI or BglII had two large hybridization fragments not different from control DNA samples. These data indicate the presence of two different 21-hydroxylase genes. Additional mapping studies revealed that the two genes had the restriction pattern of the inactive 21-hydroxylase gene. When genomic DNA that had been isolated from all members of this family and from normal subjects was hybridized with the human C4 cDNA probe, the restriction fragment hybridization patterns for all four endonuclease digests were similar in the two groups. Hence, our results suggest that the 21-hydroxylase deficiency of our patients is due to conversion of the active 21-hydroxylase gene to the inactive gene. This gene conversion was associated with absence of functional C4B protein, without any detectable alterations in the restriction fragment pattern of the C4 genes.

Original languageEnglish (US)
Pages (from-to)995-1002
Number of pages8
JournalJournal of Clinical Endocrinology and Metabolism
Volume62
Issue number5
StatePublished - 1986

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Steroid 21-Hydroxylase
Congenital Adrenal Hyperplasia
Gene Conversion
Complement System Proteins
Salts
Genes
DNA
Lymphocytes
Complement C4
Siblings
DNA Restriction Enzymes
Southern Blotting
Antigens
Adrenocorticotropic Hormone
Complementary DNA
Endonucleases
varespladib methyl
Fathers
Alleles
Mothers

ASJC Scopus subject areas

  • Biochemistry
  • Endocrinology, Diabetes and Metabolism

Cite this

Gene conversion in salt-losing congenital adrenal hyperplasia with absent complement C4B protein. / Donohoue, P. A.; van Dop, C.; McLean, R. H.; White, P. C.; Jospe, N.; Migeon, C. J.

In: Journal of Clinical Endocrinology and Metabolism, Vol. 62, No. 5, 1986, p. 995-1002.

Research output: Contribution to journalArticle

Donohoue, P. A. ; van Dop, C. ; McLean, R. H. ; White, P. C. ; Jospe, N. ; Migeon, C. J. / Gene conversion in salt-losing congenital adrenal hyperplasia with absent complement C4B protein. In: Journal of Clinical Endocrinology and Metabolism. 1986 ; Vol. 62, No. 5. pp. 995-1002.
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AU - Donohoue, P. A.

AU - van Dop, C.

AU - McLean, R. H.

AU - White, P. C.

AU - Jospe, N.

AU - Migeon, C. J.

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N2 - Two of four siblings expressed the salt-losing form of congenital adrenal hyperplasia due to 21-hydroxylase deficiency (CAH) and had identical human lymphocyte antigen (HLA) and complement C4 (fourth component of complement) types (HLA-A3, C4, B35, C4A3, C4BQO, DR1/A2, C-,B18, C4A3, C4BQO,DR6). The father and one unaffected sibling were heterozygous carriers of CAH, as determined by a 30-min iv ACTH stimulation test and HLA typing. In addition, the iv ACTH stimulation test revealed that the mother and the other unaffected sibling also carried an allele for an attenuated form of CAH. Restriction endonuclease digests of genomic DNA obtained from members of this family and from normal unrelated subjects were hybridized with cDNA probes encoding human 21-hydroxylase and C4. With the 21-hydroxylase probe, Southern blots prepared from control DNA samples revealed two major restriction fragments in each of four restriction endonuclease digests; TaqI produced major bands at 3.7 and 3.2 kilobases (kb), KpnI at 4.0 and 2.9 kb, EcoRI at 18 and 13 kb, and BglII at 15 and 12.5 kb. Southern blots prepared from DNA of the two patients lacked the 3.7-kb TaqI and 2.9-kb KpnI fragments, but had increased hybridization intensity (relative to control DNA samples) in the 3.2-kb TaqI and 4.0-kb KpnI fragments. By contrast, blots with EcoRI or BglII had two large hybridization fragments not different from control DNA samples. These data indicate the presence of two different 21-hydroxylase genes. Additional mapping studies revealed that the two genes had the restriction pattern of the inactive 21-hydroxylase gene. When genomic DNA that had been isolated from all members of this family and from normal subjects was hybridized with the human C4 cDNA probe, the restriction fragment hybridization patterns for all four endonuclease digests were similar in the two groups. Hence, our results suggest that the 21-hydroxylase deficiency of our patients is due to conversion of the active 21-hydroxylase gene to the inactive gene. This gene conversion was associated with absence of functional C4B protein, without any detectable alterations in the restriction fragment pattern of the C4 genes.

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