A Gly565-->Ala substitution in human erythroid band 3 accounts for the Wu blood group polymorphism.
BACKGROUND
Reports published in 1976 and 1980 described the low-incidence red cell antigen Wu. Distinction of Wu from all other known low-incidence antigens and from the ABO, Rh, Lutheran, Duffy, Kidd, P, and X-linked blood group systems allowed Wu to be placed in the International Society of Blood Transfusion's 700 series, designated as 700013. Recently, a blood donor apparently homozygous for Wu has been identified. This report documents the serologic and molecular findings of samples from this individual and the members of his family.


METHODS
Blood samples from 26 members of a kindred of Dutch descent segregating for Wu were collected and analyzed. Red cells were subjected to titration and enzymatic tests, while DNA was analyzed by polyacrylamide gel electrophoresis for single-strand conformational polymorphism (SSCP) and nucleotide differences by DNA sequencing.


RESULTS
Serologic investigations conducted on red cells of the propositus and two of his siblings consistently revealed higher titers with various sera containing anti-Wu than did cells from their parents or children. Treatment of intact red cells with alpha-chymotrypsin completely abolished Wu recognition. Because erythroid band 3 is cleaved by alpha-chymotrypsin, the possible relationship between Wu and AE1 (the gene controlling erythroid band 3 expression) was investigated by molecular methods. SSCP analysis of DNA revealed that all Wu+ family members exhibited a mobility shift in exon 14 of AE1. The nature of the SSCP was defined by DNA sequencing as a G-->C mutation that resulted in a Gly565-->Ala substitution in human erythroid band 3.


CONCLUSIONS
Three members of the kindred are homozygous for the low-incidence red cell antigen Wu. A G-->C mutation in AE1 gives rise to a Gly565-->Ala substitution in band 3, thereby accounting for the Wu red cell polymorphism. In light of these findings, the International Society of Blood Transfusion Working Party has provisionally assigned Wu to the Diego blood group system (designated 010009 or D19).
Department of Pediatrics, Faculty of Medicine, University of Manitoba, Winnipeg, Canada.
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Zelinski
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McManus
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Punter
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Moulds
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Coghlan
D000409:Alanine / Q000235:genetics; D001457:Anion Exchange Protein 1, Erythrocyte / Q000235:genetics*; D001789:Blood Group Antigens / Q000235:genetics*; D004591:Electrophoresis, Polyacrylamide Gel; D005260:Female; D005998:Glycine / Q000235:genetics; D006720:Homozygote; D006801:Humans; D007519:Isoantigens / Q000235:genetics*; D008297:Male; D010375:Pedigree; D017354:Point Mutation; D011110:Polymorphism, Genetic; D018807:Polymorphism, Single-Stranded Conformational; D017422:Sequence Analysis, DNA
D001457:Anion Exchange Protein 1, Erythrocyte; D001789:Blood Group Antigens; D007519:Isoantigens; D000409:Alanine; D005998:Glycine