title: Amino acid substitutions in human erythroid protein band 3 account for the low-incidence antigens NFLD and BOW.
abstract: BACKGROUND
  The low-incidence red cell antigens NFLD (700.37) and BOW (700.46) were first described in 1984 and 1988, respectively. Recent investigations showed that antigens of the Diego blood group system (including a number of low-incidence antigens) are coded by SLC4A1 (solute carrier family 4, anion exchanger member 1 gene). Among these newly characterized Diego system antigens is Wu (designated DI9). Because a serologic relationship among Wu, NFLD, and BOW has been established, a series of genetic and molecular investigations of SLC4A1 in relation to NFLD and BOW were undertaken.
  
  
  METHODS
  By the use of exon-specific primers, single-strand conformational polymorphism (SSCP) analysis of SLC4A1 was performed on DNA isolated from an NFLD+ person from Japan, from the members of a Canadian kindred segregating for NFLD, and from two unrelated BOW+ persons. Exons displaying SSCPs were subjected to genetic linkage analysis (for NFLD only) and DNA sequencing.
  
  
  RESULTS
  SSCPs in DNA amplified from exons 12 and 14 of SLC4A1 were observed for all NFLD+ subjects. Linkage between each of these polymorphisms and NFLD was established with peak lods = 4.82 at theta = 0.00 for combined paternal and maternal meiosis. DNA sequencing of exons 12 and 14 of SLC4A1 from NFLD+ persons identified A-->T and C-->G mutations that underlie Glu429Asp and Pro561Ala substitutions in human erythroid band 3 protein (band 3). DNA from the two unrelated BOW+ persons only exhibited an SSCP in exon 14 of SLC4A1. Subsequent DNA sequencing revealed a C-->T mutation that accounts for a Pro561Ser substitution in band 3.
  
  
  CONCLUSIONS
  SLC4A1 codes for the low-incidence red cell antigens NFLD and BOW. In light of these findings, both antigens have been assigned to the Diego blood group system.
authors.affiliation: Rh Laboratory, Departments of Pediatrics and Child Health and of Human Genetics, Faculty of Medicine, University of Manitoba, Winnipeg, Manitoba, Canada.
authors.forename: K
authors.initials: K
authors.lastname: McManus
authors.forename: J
authors.initials: J
authors.lastname: Pongoski
authors.forename: G
authors.initials: G
authors.lastname: Coghlan
authors.forename: T
authors.initials: T
authors.lastname: Zelinski
mesh_terms: D019943:Amino Acid Substitution; D001457:Anion Exchange Protein 1, Erythrocyte / Q000737:chemistry / Q000235:genetics*; D001789:Blood Group Antigens / Q000235:genetics / Q000276:immunology*; D054421:Chemokine CCL21; D019742:Chemokines, CC / Q000235:genetics; D005091:Exons / Q000235:genetics; D008040:Genetic Linkage; D006801:Humans; D007519:Isoantigens / Q000235:genetics; D017354:Point Mutation; D018807:Polymorphism, Single-Stranded Conformational; D017422:Sequence Analysis, DNA
chemical_list: D001457:Anion Exchange Protein 1, Erythrocyte; D001789:Blood Group Antigens; C515478:CCL21 protein, human; D054421:Chemokine CCL21; D019742:Chemokines, CC; D007519:Isoantigens