European Journal of Pediatrics

, Volume 152, Issue 6, pp 469–472 | Cite as

A newly recognized point mutation in the cytochrome b558 heavy chain gene replacing alanine57 by glutamic acid, in a patient with cytochrome b positive X-linked chronic granulomatous disease

  • T. Ariga
  • Y. Sakiyama
  • K. Tomizawa
  • S. Imajoh-Ohmi
  • S. Kanegasaki
  • S. Matsumoto
Hematology/Oncology

Abstract

Molecular genetic analysis was performed in a patient with cytochrome b positive X-linked chronic granulomatous disease. A previous Southern blot study, using a cytochrome b heavy chain cDNA as probe, revealed a Pst I restriction fragment pattern for the cytochrome b heavy chain gene (CYBB) different to that of normal individuals. Since restriction length polymorphism with Pst I has never been observed in control individuals and no abnormal restriction fragment patterns in the patient's CYBB was detected with seven other enzymes used, we focussed on the single Pst I site in the CYBB cDNA as being the only mutation site responsible for his disease. A fragment of the patient's cDNA which included the Pst I site was amplified by reverse polymerase chain reaction, and loss of the Pst I site in the fragment was confirmed by incubation with Pst I. Subsequent sequence analysis of the fragment revealed a point mutation in the Pst I site (cytosine to adenine), substituting glutamic acid for alanine at position 57.

Key words

X-linked chronic granulomatous disease Mutant cytochrome b heavy chain Point mutation Molecular genetic analysis 

Abbreviations

CGD

chronic granulomatous disease

CYBB

cytochrome b heavy chain gene

EBV

Epstein-Barr virus

phox

phagocytic oxidase

gp91-phox

91-kD glycoprotein

p22-phox

22-kD polypepetide

p47-phox

47-kD cytoplasmic oxidase component

p67-phox

67-kD cytoplasmic oxidase component

PCR

polymerase chain reaction

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. 1.
    Ariga T, Igarashi T, Ramesh N, Parad R, Cicardi M, Davis AEIII (1989) Type I C1 inhibitor deficiency with a small messenger RNA resulting from deletion of one exon. J Clin Invest 83:1888–1893PubMedGoogle Scholar
  2. 2.
    Ariga T, Nakanishi M, Tomizawa K, Imajoh-Ohmi S, Kanegasaki S, Sakiyama S, Matsumoto S (1992) Genetic heterogeneity in patients with X-linked recessive chronic granulomatous disease. Pediatr Res 31:516–519PubMedGoogle Scholar
  3. 3.
    Battat L, Francke U (1989) Nsi I RFLP at the X-linked chronic granulomatous disease locus (CYBB). Nucleic Acids Res 17: 3619PubMedGoogle Scholar
  4. 4.
    Bolsher BGJM, Boer M, Klein A, Weening RS, Roos D (1991) Point mutations in the β-subunit of cytochrome b558 leading to X-linked chronic granulomatous disease. Blood 77: 248–287Google Scholar
  5. 5.
    Dinauer MC, Curnutte JT, Rosen H, Orkin SH (1989) A missense mutation in the neutrophil cytochrome b heavy chain in cytochrome-positive X-linked chronic granulomatous disease. J Clin Invest 84:2012–2016PubMedGoogle Scholar
  6. 6.
    Imajoh-Ohmi S, Tokita K, Ochiai H, Nakamura M, Kanegasaki S (1992) Topology of cytochrome b558 in neutrophil membrane analyzed by anti-peptide antibodies and proteolysis. J Biol Chem 267:180–184PubMedGoogle Scholar
  7. 7.
    Orkin SH (1989) Molecular genetics of chronic granulomatous disease. Ann Rev Immunol 7:277–307Google Scholar
  8. 8.
    Rotrosen D, Kleinberg ME, Nunoi H, Leto T, Gallin JI, Malech HL (1990) Evidence for a functional cytoplasmic domain of phagocyte oxidase cytochrome b558. J Biol Chem 265:8745–8750PubMedGoogle Scholar
  9. 9.
    Saiki R, Gelfand D, Stoffel S, Scharf S, Higuchi R, Horn G, Mullis K, Ehrlich H (1988) Primer-directed enzymatic amplification of DNA with a thermostable DNA polymerase. Science 239:487–494PubMedGoogle Scholar
  10. 10.
    Sanger F, Nicklen S, Coulson RA (1987) DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci USA 74: 5463–5467Google Scholar
  11. 11.
    Segal AW (1987) Absence of both cytochrome b-245 subunits from neutrophils in X-linked chronic granulomatous disease. Nature 326:88–91PubMedGoogle Scholar
  12. 12.
    Shapiro BL, Newburger PE, Klempner MS, Dinauer MC (1991) Chronic granulomatous disease in a 69-year old man. N Engl J Med 325:1786–1790PubMedGoogle Scholar
  13. 13.
    Shurin SB, Cohen HJ, Whitin JC, Newburger PE (1983) Impaired granulocyte superoxide production and prolongation of the respiratory burst due to a low affinity NADPH-dependent oxidase. Blood 62:564–571PubMedGoogle Scholar
  14. 14.
    Skalnik DG, Strauss EC, Orkin SH (1991) CCAAT displacement protein as a repressor of the myelomonocytic-specific gp91-phox gene promoter. J Biol Chem 266:16736–16744PubMedGoogle Scholar
  15. 15.
    Smith RB, Curnutte JT (1991) Molecular basis of chronic granulomatous disease. Blood 77:673–686PubMedGoogle Scholar
  16. 16.
    Tomizawa K, Sakiyama Y, Matsumoto S, Nakamura M (1991) The expression of 22-kD subunit of cytochrome b558 in patients with X-linked chronic granulomatous disease. Eur J Haematol 46:52–54PubMedGoogle Scholar
  17. 17.
    Whitehead AS, Woods DE, Fleishnic JE, Yunis EJ, Katz AJ, Gerald PS, Alper CA, Colten HR (1984) DNA polymorphism of the C4 genes: a new marker for analysis of the major histocompatibility complex. N Engl J Med 310:88–91PubMedGoogle Scholar

Copyright information

© Springer-Verlag 1993

Authors and Affiliations

  • T. Ariga
    • 1
  • Y. Sakiyama
    • 1
  • K. Tomizawa
    • 1
  • S. Imajoh-Ohmi
    • 2
  • S. Kanegasaki
    • 2
  • S. Matsumoto
    • 1
  1. 1.Department of PaediatricsHokkaido University School of MedicineSapporoJapan
  2. 2.Institute of Medical ScienceUniversity of TokyoTokyoJapan

Personalised recommendations