Ankyrin–1 mutations are a major cause of dominant and recessive hereditary spherocytosis

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Hereditary spherocytosis (HS) is the most common inherited haemolytic anaemia in Northern Europeans. The primary molecular defects reside in the red blood cell (RBC) membrane, particularly in proteins that link the membrane skeleton to the overlying lipid bilayer and its integral membrane constituents1. Ankyrin-1 is the predominant linker molecule. It attaches spectrin, the major skeletal protein, to the cytoplasmic domain of band 3, the RBC anion exchanger. Two-thirds of patients with HS have combined spectrin and ankyrin-1 deficiency2,3; deficiency of band 3 occurs in about 15 to 20% (ref. 1). These data suggest that ankyrin-1 or band 3 defects may be common in HS. To test this we screened all 42 coding exons plus the 5′ untranslated/promoter region of ankyrin-1 and the 19 coding exons of band 3 in 46 HS families. Twelve ankyrin-1 mutations and five band 3 mutations were identified. Missense mutations and a mutation in the putative ankyrin-1 promoter were common in recessive HS. In contrast, ankyrin-1 and band 3 frameshift and nonsense null mutations prevailed in dominant HS. Increased accu-mulation of the normal protein product partially compensated for the ankyrin-1 or band 3 defects in some of these null mutations. Our findings indi-cate that ankyrin-1 mutations are a major cause of dominant and recessive HS (35 to 65%), that band 3 mutations are less common (15 to 25%), and that the severity of HS is modified by factors other than the primary gene defect.

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  1. 1

    Lux, S.E. & Palek, J. Disorders of the red cell membrane, in Blood: Principles and Practice of Hematology (eds Handin, R.I., Lux, S.E. & Stossel, T.R) 1701–1818 (J.B. Lippincott, Philadelphia, 1995).

  2. 2

    Pekrun, A., Eber, S.W., Kuhlmey, A. & Schröter, W. Combined ankyrin and spectrin deficiency in hereditary spherocytosis. Ann. Hematol. 67, 89–93 (1993).

  3. 3

    Savvides, P., Shalev, O., John, K.M. & Lux, S.E. Combined spectrin and ankyrin deficiency is common in autosomal dominant hereditary spherocytosis. Blood 82, 2953–2960 (1993).

  4. 4

    Lux, S.E., John, K.M. & Bennett, V. Analysis of cDNA for human erythrocyte ankyrin indicates a repeated structure with homology to tissue-differentiation and cell-cycle control proteins. Nature 344, 36–43 (1990).

  5. 5

    Lux, S.E., John, K.M., Kopito, R.R. & Lodish, H.F. Cloning and characterization of band 3, the human erythrocyte anion-exchange protein (AE1). Proc. Natl. Acad. Sci. USA 86, 9088–9093 (1989).

  6. 6

    Beaudet, A.L. & Tsui, L.C. A suggested nomenclature for designating mutations.Hum. Mutat. 2, 245–248 (1993).

  7. 7

    Hanspal, M. et al. Molecular basis of spectrin and ankyrin deficiencies in severe hereditary spherocytosis: evidence implicating a primary defect of ankyrin. Blood 77, 165–173 (1991).

  8. 8

    Lux, S.E. et al. Hereditary spherocytosis associated with deletion of the human erythrocyte ankyrin gene on chromosome 8. Nature. 345, 736–739 (1990).

  9. 9

    Costa, F.F. et al. Linkage of dominant hereditary spherocytosis to the gene for the erythrocyte membrane-skeleton protein ankyrin. New Engl.J. Med. 323, 1046–1050 (1990).

  10. 10

    Jarolim, P., Rubin, H.L., Barbec, V. & Palek, J. A nonsense mutation 1669 Glu→Ter within the regulatory domain of human erythroid ankyrin leads to a selective deficiency of the major ankyrin isoform (band 2.1) and a phenotype of autosomal dominant hereditary spherocytosis. J. Clin. Invest. 95, 941–947 (1995).

  11. 11

    Gallagher, P.G., Romana, M., Tse, W.T., Eber, S., Lux, S.E. & Forget, B.G. Erythroid-specific expression of the human erythrocyte ankyrin gene is mediated by a promoter that displays features found in both housekeeping and erythroid gene promoters. Blood 84, 361a (1994).

  12. 12

    Eber, S.W., Pekrun, A., Reinhardt, D., Schroter, W. & Lux, S.E. Hereditary spherocytosis with ankyrin Walsrode, a variant ankyrin with decreased affinity for band 3. Blood. 84, 362a (1994).

  13. 13

    Gallagher, P.G., Tse, W.T., Scarpa, A.L., Lux, S.E. & Forget, B.G. Large number of alternatively spliced isoforms of the regulatory region of human erythrocyte ankyrin. Trans. Assoc. Am. Phys. 105, 268–277 (1992).

  14. 14

    Agre, P., Asimos, A., Casella, J.F. & McMillan, D. Inheritance pattern and clinical response to splenectomy as a reflection of erythrocyte spectrin deficiency in hereditary spherocytosis. NewEngl. J. Med. 315, 1579–1583 (1986).

  15. 15

    Wilmotte, R. et al. Low expression allele αLELY of red cell spectrin is associated with mutations in exon 40 (αv/41 polymorphism) and intron 45 and with partial skipping of exon 46. J. Clin. Invest. 91, 2091–2096 (1993).

  16. 16

    Sheffield, V.C., Beck, J.S., Kwitek, A.E., Sandstrom, D.W. & Stone, E.M. The sensitivity of single-strand conformation polymorphism analysis for the detection of single base substitutions. Genomics 16, 325–332 (1993).

  17. 17

    Liu, Q. & Sommer, S.S. Parameters affecting the sensitivities of dideoxy fingerprinting and SSCP. PCR Meth.Applic. 4, 97–108 (1994).

  18. 18

    Hayashi, K. & Yandell, D.W. How sensitive is PCR SSCP? Hum. Mutat. 2, 338–346 (1993).

  19. 19

    Michaud, J. et al. Strand-separating conformational polymorphism analysis: efficacy of detection of point mutations in the human ornithine δ-aminotransferase gene. Genomics. 13, 389–394 (1992).

  20. 20

    Sarkar, G., Yoon, H-S. & Sommer, S.S. Screening for mutations by RNA single-strand conformation polymorphism (rSSCP): comparison with DNA-SSCR Nucl. Acids Res. 20, 871–678 (1992).

  21. 21

    Jarolim, P., Rubin, H.L., Brabec, V. & Palek, J. Comparison of the ankyrin (AC)n microsatellites in genomic DMA and mRNA reveals absence of one ankyrin mRNA allele in 20% of patients with hereditary spherocytosis. Blood 85, 3278–3282 (1995).

  22. 22

    Mclntosh, I., Hamosh, A. & Dietz, H.C. Nonsense mutations and diminished mRNA levels. Nature Genet. 4, 219 (1993).

  23. 23

    Jarolim, P. et al. Duplication of 10 nucleotides in the erythroid band 3 (AE1) gene in a kindred with hereditary spherocytosis and band 3 protein deficiency (Band 3PRAGUE). J. Clin. Invest. 93, 121–130 (1994).

  24. 24

    Jarolim, P. et al. Mutations of conserved arginines in the membrane domain of erythroid band 3 lead to a decrease in membrane-associated band 3 and to the phenotype of hereditary spherocytosis. Blood 85, 634–640 (1995).

  25. 25

    Jarolim, P., Murray, J., Rubin, K.L., Palek, J. & Hemolytic Anemia Study Group. Molecular characterization of hereditary spherocytosis with band 3 deficiency. Blood 84, 362a (1994).

  26. 26

    Dalbey, R.E. Positively charged residues are important determinants of membrane protein topology. Trends Biol. Sci. 15, 253–257 (1990).

  27. 27

    Hassoun, H. et al. Characterization of the underlying molecular defect in hereditary spherocytosis associated with spectrin deficiency. Blood 86, 467a(1995).

  28. 28

    Wichterle, H. et al. Two recessively inherited defects of α-spectrin underlie a severe spectrin-deficient spherocytic hemolytic anemia. Blood. 86, 468a (1995).

  29. 29

    Peters, L.L. et al. Targeted disruption of the band 3 gene in mice causes severe hemolytic anemia despite a nearly normal membrane skeletal assembly. Blood 86, 124a(1995).

  30. 30

    Eber, S.W., Armbrust, R. & Schröter, W. Variable clinical severity of hereditary spherocytosis: relation to erythrocytic spectrin concentration, osmotic fragility and autohemolysis. J. Pediatr. 177, 409–416 (1990).

  31. 31

    Bell, G.I., Karam, J.H. & Rutter, W.J. Polymorphic DNA region adjacent to the 5′ end of the human insulin gene. Proc. Nati Acad. Sci. USA 78, 5759–5763 (1981).

  32. 32

    Laemmli, U.K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227, 680–685 (1970).

  33. 33

    Ausubel, F.M. et al. in Current Protocols in Molecular Biology (John Wiley & Sons, New York, 1995).

  34. 34

    Soto, D. & Sukumar, S. Improved detection of mutations in the p53 gene in human tumors as single-stranded conformation polymorphism and double-stranded heteroduplex DNA. PCP, Meth. Appl. 2, 96–98 (1992).

  35. 35

    Goosens, M. & Kan, Y.W. DNA analysis in the diagnosis of hemoglobin disorders. Meth. Enzymol. 76, 805–817 (1981).

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