Skip to main content
NCBI home page
Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1982 Jan;79(1):137–141. doi: 10.1073/pnas.79.1.137

Nonrandom association of polymorphic restriction sites in the β-globin gene cluster

Stylianos E Antonarakis *, Corinne D Boehm *, Patricia J V Giardina †, Haig H Kazazian Jr *
PMCID: PMC345677  PMID: 6275383

Abstract

By using probes for ε-, Ψβ1-, and β-globin genes, we found four additional polymorphic restriction sites that have frequencies >0.1 in persons of Mediterranean area origin, Asian Indians, and American Blacks. Three of these (HincII sites) and the two previously described polymorphic HindIII sites [one in intervening sequence (IVS) II of each γ-globin gene] are distributed over 32 kilobases (kb) of DNA located 5′ to the δ-globin gene. This region of DNA comprises two-thirds of the β-globin gene cluster. Since each of these five polymorphic sites can be present (+) or absent (-), in theory there exist 32 possible combinations of sites (haplotypes). However, in Italians, Greeks, Indians, and Turks, 3 of the 32 haplotypes, (+----), (-+-++), and (-++-+), account for 92% of 89 βA chromosomes examined. The observed frequencies for these haplotypes are 0.64, 0.15, and 0.13 in the populations studied, in contrast to expected frequencies (based on the observed gene frequencies at each of the five sites) of 0.20, 0.006, and 0.005, respectively. In American Blacks, a fourth haplotype, (----+), which is rare in non-Black populations, has a frequency of 0.37 in contrast to its expected frequency of 0.05. These results suggest a nonrandom association of DNA sequences over 32 kb 5′ to the δ-globin gene in all populations studied. Two other polymorphic sites 3′ to the δ gene (the newly discovered Ava II site in IVS II of the β-globin gene and the BamHI site 3′ to it) are nonrandomly associated with each other but randomly distributed with respect to the above haplotypes. This suggests that randomization of sequences has occurred within 12 kb of DNA between these two nonrandomly associated sequence clusters. Nonrandom association of polymorphic restriction sites has practical consequences in that it limits the usefulness of these additional HincII sites for prenatal diagnosis of hemoglobinopathies by linkage analysis. These sites provide little additional information for detection of β-thalassemia, while the polymorphic Ava II site, which lies outside the nonrandomly associated sequences 5′ to the δ gene, improves the test applicability from 52% to 70% of couples at risk.

Keywords: DNA polymorphism, hemoglobinopathies, population haplotype frequencies, prenatal diagnosis

Full text

PDF
137

Images in this article

138Image
on p.138

Selected References

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

  1. Efstratiadis A., Posakony J. W., Maniatis T., Lawn R. M., O'Connell C., Spritz R. A., DeRiel J. K., Forget B. G., Weissman S. M., Slightom J. L. The structure and evolution of the human beta-globin gene family. Cell. 1980 Oct;21(3):653–668. doi: 10.1016/0092-8674(80)90429-8. [DOI] [PubMed] [Google Scholar]
  2. Fritsch E. F., Lawn R. M., Maniatis T. Molecular cloning and characterization of the human beta-like globin gene cluster. Cell. 1980 Apr;19(4):959–972. doi: 10.1016/0092-8674(80)90087-2. [DOI] [PubMed] [Google Scholar]
  3. Giblett E. R. Genetic polymorphisms in human blood. Annu Rev Genet. 1977;11:13–28. doi: 10.1146/annurev.ge.11.120177.000305. [DOI] [PubMed] [Google Scholar]
  4. Harris H., Hopkinson D. A. Average heterozygosity per locus in man: an estimate based on the incidence of enzyme polymorphisms. Ann Hum Genet. 1972 Jul;36(1):9–20. doi: 10.1111/j.1469-1809.1972.tb00578.x. [DOI] [PubMed] [Google Scholar]
  5. Jeffreys A. J. DNA sequence variants in the G gamma-, A gamma-, delta- and beta-globin genes of man. Cell. 1979 Sep;18(1):1–10. doi: 10.1016/0092-8674(79)90348-9. [DOI] [PubMed] [Google Scholar]
  6. Jeffreys A. J., Flavell R. A. A physical map of the DNA regions flanking the rabbit beta-globin gene. Cell. 1977 Oct;12(2):429–439. doi: 10.1016/0092-8674(77)90119-2. [DOI] [PubMed] [Google Scholar]
  7. Kan Y. W., Dozy A. M. Antenatal diagnosis of sickle-cell anaemia by D.N.A. analysis of amniotic-fluid cells. Lancet. 1978 Oct 28;2(8096):910–912. doi: 10.1016/s0140-6736(78)91629-x. [DOI] [PubMed] [Google Scholar]
  8. Kan Y. W., Dozy A. M. Polymorphism of DNA sequence adjacent to human beta-globin structural gene: relationship to sickle mutation. Proc Natl Acad Sci U S A. 1978 Nov;75(11):5631–5635. doi: 10.1073/pnas.75.11.5631. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Kan Y. W., Lee K. Y., Furbetta M., Angius A., Cao A. Polymorphism of DNA sequence in the beta-globin gene region. Application to prenatal diagnosis of beta 0 thalassemia in Sardinia. N Engl J Med. 1980 Jan 24;302(4):185–188. doi: 10.1056/NEJM198001243020401. [DOI] [PubMed] [Google Scholar]
  10. Kazazian H. H., Jr, Phillips J. A., 3rd, Boehm C. D., Vik T. A., Mahoney M. J., Ritchey A. K. Prenatal diagnosis of beta-thalassemias by amniocentesis: linkage analysis using multiple polymorphic restriction endonuclease sites. Blood. 1980 Nov;56(5):926–930. [PubMed] [Google Scholar]
  11. Kunkel L. M., Smith K. D., Boyer S. H., Borgaonkar D. S., Wachtel S. S., Miller O. J., Breg W. R., Jones H. W., Jr, Rary J. M. Analysis of human Y-chromosome-specific reiterated DNA in chromosome variants. Proc Natl Acad Sci U S A. 1977 Mar;74(3):1245–1249. doi: 10.1073/pnas.74.3.1245. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Kurnit D. M., Hoehn H. Prenatal diagnosis of human genome variation. Annu Rev Genet. 1979;13:235–258. doi: 10.1146/annurev.ge.13.120179.001315. [DOI] [PubMed] [Google Scholar]
  13. Little P. F., Annison G., Darling S., Williamson R., Camba L., Modell B. Model for antenatal diagnosis of beta-thalassaemia and other monogenic disorders by molecular analysis of linked DNA polymorphisms. Nature. 1980 May 15;285(5761):144–147. doi: 10.1038/285144a0. [DOI] [PubMed] [Google Scholar]
  14. Orkin S. H., Goff S. C. Nonsense and frameshift mutations in beta 0-thalassemia detected in cloned beta-globin genes. J Biol Chem. 1981 Oct 10;256(19):9782–9784. [PubMed] [Google Scholar]
  15. Panny S. R., Scott A. F., Smith K. D., Phillips J. A., 3rd, Kazazian H. H., Jr, Talbot C. C., Jr, Boehm C. D. Population heterogeneity of the Hpa I restriction site associated with the beta globin gene: implications for prenatal diagnosis. Am J Hum Genet. 1981 Jan;33(1):25–35. [PMC free article] [PubMed] [Google Scholar]
  16. Phillips J. A., 3rd, Panny S. R., Kazazian H. H., Jr, Boehm C. D., Scott A. F., Smith K. D. Prenatal diagnosis of sickle cell anemia by restriction and endonuclease analysis: HindIII polymorphisms in gamma-globin genes extend test applicability. Proc Natl Acad Sci U S A. 1980 May;77(5):2853–2856. doi: 10.1073/pnas.77.5.2853. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Proudfoot N. J., Baralle F. E. Molecular cloning of human epsilon-globin gene. Proc Natl Acad Sci U S A. 1979 Nov;76(11):5435–5439. doi: 10.1073/pnas.76.11.5435. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Reed T. E. Caucasian genes in American Negroes. Science. 1969 Aug 22;165(3895):762–768. doi: 10.1126/science.165.3895.762. [DOI] [PubMed] [Google Scholar]
  19. Schachat F. H., Hogness D. S. Repetitive sequences in isolated Thomas circles from Drosophila melanogaster. Cold Spring Harb Symp Quant Biol. 1974;38:371–381. doi: 10.1101/sqb.1974.038.01.040. [DOI] [PubMed] [Google Scholar]
  20. Scott A. F., Phillips J. A., 3rd, Migeon B. R. DNA restriction endonuclease analysis for localization of human beta- and delta-globin genes on chromosome 11. Proc Natl Acad Sci U S A. 1979 Sep;76(9):4563–4565. doi: 10.1073/pnas.76.9.4563. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Southern E. M. Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol. 1975 Nov 5;98(3):503–517. doi: 10.1016/s0022-2836(75)80083-0. [DOI] [PubMed] [Google Scholar]
  22. Tuan D., Biro P. A., deRiel J. K., Lazarus H., Forget B. G. Restriction endonuclease mapping of the human gamma globin gene loci. Nucleic Acids Res. 1979 Jun 11;6(7):2519–2544. doi: 10.1093/nar/6.7.2519. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Wilson J. T., Wilson L. B., deRiel J. K., Villa-komaroff L., Efstratiadis A., Forget B. G., Weissman S. M. Insertion of synthetic copies of human globin genes into bacterial plasmids. Nucleic Acids Res. 1978 Feb;5(2):563–581. doi: 10.1093/nar/5.2.563. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Proceedings of the National Academy of Sciences of the United States of America are provided here courtesy of National Academy of Sciences

RESOURCES