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Cited by 569Sexual Differentiation and DisordersAdrenal Hormones and DisordersHormonal Regulation and Hypertension

Structure of human steroid 21-hydroxylase genes.

Open access
White, P C, New, M I, Dupont, B·1986·Proceedings of the National Academy of Sciences
DOI 10.1073/pnas.83.14.5111PMC PMC323900
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Abstract

We have determined the structure of cDNA and two genomic genes encoding steroid 21-hydroxylase [21-OHase; steroid 21-monooxygenase; steroid, hydrogen-donor:oxygen oxidoreductase (21-hydroxylating); EC 1.14.99.10]. If this cytochrome P-450 enzyme is defective, cortisol cannot be synthesized, resulting in congenital adrenal hyperplasia. The cDNA encoding this enzyme is 2.0 kilobases long, and the encoded protein is predicted to contain 494 amino acid residues with a molecular weight of 55,000. This enzyme is at most 28% homologous to other P-450 enzymes that have been studied. The 21-OHase genomic genes, which are located in the HLA major histocompatibility complex on chromosome 6, each contain 10 exons. This structure is distinct from other characterized P-450 genes, which contain 7 or 9 exons. Studies of individuals with homozygous deletions of the 21-OHase A or B genes suggest that only the B gene encodes an active enzyme. This is confirmed by the finding that the A gene has an 8-base deletion within codons 110-112, resulting in a frameshift that brings a stop codon into the reading frame at codon 130. A second frameshift and a nonsense mutation occur downstream. In contrast, the sequence of the exons of the B gene is identical to the cDNA sequence. The 21-OHase A gene is, therefore, a pseudogene.

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  • 21-hydroxylase deficiency is one of the most technically challenging monogenic conditions for molecular diagnosis because of the complex genomic architecture of the CYP21A2 locus and the extensive homology between the functional gene and its pseudogene.

    “Congenital adrenal hyperplasia (CAH) due to 21-hydroxylase deficiency is one of the most technically challenging monogenic conditions for molecular diagnosis, owing to the complex genomic architecture of the CYP21A2 locus and the extensive homology between the functional gene and its pseudogene.”

    From Evolution of molecular diagnostic strategies for 21-hydroxylase deficiency: from classical methods to advanced genomic techniques (2026) · cites this paper as [9]

    The paper’s Abstract identifies an active 21-hydroxylase B gene and an A pseudogene, and its gene-structure section reports 98% exon and 96% intron homology between them (page not available in our copy). These findings support the claimed genomic architecture and extensive homology. The paper does not characterize molecular diagnosis of this condition as one of the most technically challenging among monogenic conditions. (LLM, conf=0.86)

    Experimental — accuracy not yet measured

  • Functional CYP21A2 is adjacent to C4B, extends over approximately 3.4 kb, and encodes a 495-amino-acid cytochrome P450 enzyme, whereas CYP21A1P is adjacent to C4A and is a pseudogene containing multiple inactivating pathogenic variants.

    “The functional gene CYP21A2, located adjacent to C4B, extends over approximately 3.4 kb and encodes a cytochrome P450 enzyme comprising 495 amino acids, whereas CYP21A1P, located adjacent to C4A, is a pseudogene containing multiple inactivating pathogenic variants that preclude proper protein expression (9, 10) (Figure 1).”

    From Evolution of molecular diagnostic strategies for 21-hydroxylase deficiency: from classical methods to advanced genomic techniques (2026) · cites this paper as [9]

    The Abstract and Fig. 1 place the active 21-OHase B gene beside C4B and the A pseudogene beside C4A; the genomic-gene discussion identifies multiple mutations that prevent the A gene from encoding a functional protein. But the paper predicts 494 amino acids, not 495, and places the polyadenylation signal 3.2 kb downstream of the probable transcription start rather than stating that the gene extends approximately 3.4 kb (p. 5111; “Isolation of cDNA Clones”). It also says that whether the A gene product is expressed at all had not been examined. (LLM, conf=0.90)

    Experimental — accuracy not yet measured

  • CYP21A1P is transcribed but its mRNA cannot encode a functional protein owing to at least 10 deleterious mutations including two frameshifts, a nonsense mutation, and a mutation in intron 2 that activates a cryptic splice site.

    “CYP21A1P is transcribed but its mRNA cannot encode a functional protein owing to at least 10 deleterious mutations (143,144) including two frameshifts (8 bp deletion in exon 3, 1 bp insertion in exon 7, a nonsense mutation (p.Gln318stop; Q318X) (152), and a mutation in intron 2 that activates a cryptic splice site and causes an extra 19 nucleotides to be included in the mRNA (153).”

    From Congenital Adrenal Hyperplasia—Current Insights in Pathophysiology, Diagnostics, and Management (2022) · cites this paper as [143]

    In “Isolation of cDNA Clones” (page unavailable), the paper reports that the 21-OHase A pseudogene cannot encode an active protein because of an 8-base deletion, a second frameshift, and a nonsense mutation. It does not establish at least 10 deleterious mutations; it says whether the gene is transcribed had not been examined. The passage also does not report the claimed intron-2 cryptic splice site. (LLM, conf=0.88)

    Experimental — accuracy not yet measured

  • The most important CYP21A2 transcript begins 10-11 nucleotides before the initial AUG codon.

    “The most important CYP21A2 transcript begins 10-11 nucleotides before the initial AUG codon (143).”

    From Congenital Adrenal Hyperplasia—Current Insights in Pathophysiology, Diagnostics, and Management (2022) · cites this paper as [143]

    The “Structure of the 21-OHase Genomic Genes” passage reports that transcription starts 11 bp before the ATG in the active murine gene and that the human gene has features consistent with a similar start site. It explicitly says the human transcription start had not been located. It therefore does not establish that the most important human CYP21A2 transcript starts 10–11 nucleotides before the AUG. (LLM, conf=0.87)

    Experimental — accuracy not yet measured

  • Characterization of the P450c21 protein and gene cloning show there is only one 21-hydroxylase encoded by a single functional gene on chromosome 6p21.

    “However, characterization of the P450c21 protein (355) and gene cloning show there is only one 21-hydroxylase encoded by a single functional gene on chromosome 6p21 (356 -359).”

    From The Molecular Biology, Biochemistry, and Physiology of Human Steroidogenesis and Its Disorders (2011) · cites this paper as [358]

    The Abstract reports two 21-hydroxylase genes in the chromosome 6 HLA region and identifies the A gene as a pseudogene, supporting one functional gene in the genes studied. But the discussion under “Isolation of cDNA Clones” says it was not yet known whether the A gene is nonfunctional in all individuals, so the claim’s unqualified assertion is stronger than the paper’s conclusion. (LLM, conf=0.83)

    Experimental — accuracy not yet measured

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Cites

Flagged first, then by citations

2 of the 37 references couldn’t be matched to a known paper.

Research works printed but not linked: 20

  1. [3]3. Stanbury, J. B., Wyngaarden, J. B., Fredrickson, D. S., Goldstein, J. L. & Brown, M. S. (McGraw-Hill, New York), pp. 973-1000.
  2. [6]6. White, P. C., New, M. I. & Dupont, B. (1984) Proc. Natl. Acad. Sci. USA 81, 7505-7509.
  3. [7]7. White, P. C., Grossberger, D., Onufer, B. J., Chaplin, D. D., New, M. I., Dupont, B. & Strominger, J. L. (1985) Proc. Natl. Acad. Sci. USA 82, 1089-1093.
  4. [12]12. Grunstein, M. & Hogness, D. (1975) Proc. Natl. Acad. Sci. USA 72, 3961-3965.
  5. [13]13. White, P. C., New, M. I. & Dupont, B. (1984) Proc. Natl. Acad. Sci. USA 81, 1986-1990.
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  7. [20]20. Sanger, F., Nicklen, S. & Coulson, A. R. (1977) Proc. Natl. Acad. Sci. USA 74, 5463-5467.
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  9. [24]24. Lewin, B. (1980) Gene Expression 2 (Wiley, New York), pp. 761-864.
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  11. [27]27. Mizukami, Y., Sogawa, K., Suwa, Y., Muramatsu, M. & Fujii-Kuriyama, Y. (1983) Proc. Natl. Acad. Sci. USA 80, 3958-3962.
  12. [28]28. Sogawa, K., Gotoh, O., Kawajiri, K. & Fujii-Kuriyama, Y. (1984) Proc. Natl. Acad. Sci. USA 81, 5066-5070.
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Authors
White, P. C., New, M. I., & Dupont, B.
Affiliations
Cornell University (US)
ORCID
Venue
Proceedings of the National Academy of Sciences
Publisher
National Academy of Sciences
Published
1986-07
Volume
83
Issue
14
Pages
5111-5115
ISSN
  • 0027-8424 (ISSN-L)
  • 1091-6490
DOI
10.1073/pnas.83.14.5111
PMC
PMC323900
OpenAlex
W1964331183
Semantic Scholar
ccd3407f0482a5416b4a637b0f55e07a9dd01000
Cited by
569
References
37
Citations by year
2026: 7 · 2025: 9 · 2024: 12 · 2023: 12 · 2022: 8 · 2021: 15 · 2020: 5 · 2019: 6 · 2018: 7 · 2017: 6 · 2016: 8 · 2015: 8 · 2014: 8 · 2013: 16 · 2012: 12
FWCI
9.94
Type
journal article
Language
en
Open access
Locations
  • Proceedings of the National Academy of Sciences (publishedVersion)
  • Europe PMC (PubMed Central) (publishedVersion)
  • PubMed Central (publishedVersion)
  • PubMed (publishedVersion)
Funders
  • National Cancer Institute (CA 22507, CA08478)
  • Eunice Kennedy Shriver National Institute of Child Health and Human Development (HD 00072)
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Sexual Differentiation and Disorders
  • Health Sciences › Medicine › Endocrinology, Diabetes and Metabolism › Adrenal Hormones and Disorders
  • Health Sciences › Medicine › Endocrinology, Diabetes and Metabolism › Hormonal Regulation and Hypertension
Keywords
21-hydroxylase, CYP21, cytochrome P450, congenital adrenal hyperplasia, cDNA structure, exon-intron organization, HLA complex, chromosome 6, pseudogenes, frameshift mutation, nonsense mutation, cortisol production, gene homology
MeSH terms
Amino Acid Sequence, Base Sequence, Cytochrome P-450 Enzyme System, DNA, Genes, Humans, Molecular Weight, Sequence Homology, Nucleic Acid, Steroid Hydroxylases, Steroid 21-Hydroxylase

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