title: A nonsense mutation in the erythrocyte band 3 gene associated with decreased mRNA accumulation in a kindred with dominant hereditary spherocytosis.
abstract: We studied a French kindred with typical hereditary spherocytosis (HS). Studies of erythrocytes and erythrocyte membranes from HS individuals revealed abnormal erythrocyte membrane mechanical stability as well as 15-20% deficiency of band 3, the anion transporter. Anion transport studies of red cells from two affected individuals revealed decreased sulfate flux. Nucleotide sequence of cDNA encoding the distal third of the cytoplasmic domain and the entire transmembrane domain of band 3 obtained by RT-PCR of reticulocyte RNA of an affected family member was normal. Sequence analysis of genomic DNA from an HS individual identified a nonsense mutation of the band 3 gene, Q330X, near the end of the band 3 cytoplasmic domain. This mutation was present in genomic DNA of all HS family members and absent in DNA of unaffected family members. Using an RT-PCR-based assay, a marked quantitative decrease in accumulation of the mutant band 3 RNA was detected. Thus the codon 330 nonsense mutation is responsible for the decreased accumulation of mutant band 3 RNA and the deficiency of band 3 protein in this kindred. These results have important implications for the role of band 3 defects in the membrane pathobiology of HS as well as for the techniques used in detection of HS mutations.
authors.affiliation: Department of Internal Medicine, Yale University School of Medicine, New Haven, Connecticut 06520, USA.
authors.forename: P B
authors.initials: PB
authors.lastname: Jenkins
authors.forename: G K
authors.initials: GK
authors.lastname: Abou-Alfa
authors.forename: D
authors.initials: D
authors.lastname: Dhermy
authors.forename: E
authors.initials: E
authors.lastname: Bursaux
authors.forename: C
authors.initials: C
authors.lastname: Féo
authors.forename: A L
authors.initials: AL
authors.lastname: Scarpa
authors.forename: S E
authors.initials: SE
authors.lastname: Lux
authors.forename: M
authors.initials: M
authors.lastname: Garbarz
authors.forename: B G
authors.initials: BG
authors.lastname: Forget
authors.forename: P G
authors.initials: PG
authors.lastname: Gallagher
mesh_terms: D001457:Anion Exchange Protein 1, Erythrocyte / Q000235:genetics*; D000838:Anions / Q000097:blood; D001483:Base Sequence; D001692:Biological Transport; D017931:DNA Primers / Q000737:chemistry; D004912:Erythrocytes / Q000378:metabolism; D005260:Female; D015870:Gene Expression; D005799:Genes, Dominant; D006801:Humans; D008297:Male; D008969:Molecular Sequence Data; D010375:Pedigree; D017354:Point Mutation; D012333:RNA, Messenger / Q000378:metabolism; D012156:Reticulocytes / Q000378:metabolism; D013103:Spherocytosis, Hereditary / Q000235:genetics*
chemical_list: D001457:Anion Exchange Protein 1, Erythrocyte; D000838:Anions; D017931:DNA Primers; D012333:RNA, Messenger