title: Characterization of 13 novel band 3 gene defects in hereditary spherocytosis with band 3 deficiency.
abstract: Hereditary spherocytosis (HS) is a common hemolytic anemia of variable clinical expression. Pathogenesis of HS has been associated with defects of several red cell membrane proteins including erythroid band 3. We have studied erythrocyte membrane proteins in 166 families with autosomal dominant HS. We have detected relative deficiency of band 3 in 38 kindred (23%). Band 3 deficiency was invariably associated with mild autosomal dominant spherocytosis and with the presence of pincered red cells in the peripheral blood smears of unsplenectomized patients. We hypothesized that this phenotype is caused by band 3 gene defects. Therefore, we screened band 3 DNA from these 38 kindred for single strand conformational polymorphisms (SSCP). In addition to five mutations detected previously by SSCP screening of cDNA, we detected 13 new band 3 gene mutations in 14 kindred coinherited with HS. These novel mutations consisted of two distinct subsets. The first subset included seven nonsense and frameshift mutations that were all associated with the absence of the mutant mRNA allele from reticulocyte RNA, implicating decreased production and/or stability of mutant mRNA as the cause of decreased band 3 synthesis. The second group included five substitutions of highly conserved amino acids and one in-frame deletion. These six mutations were associated with the presence of comparable levels of normal and mutant band 3 mRNA. We suggest that these mutations interfere with band 3 biosynthesis leading thus to the decreased accumulation of the mutant band 3 allele in the plasma membrane.
authors.affiliation: Department of Biomedical Research, St Elizabeth's Medical Center, Tufts University School of Medicine, Boston, MA, USA.
authors.forename: P
authors.initials: P
authors.lastname: Jarolim
authors.forename: J L
authors.initials: JL
authors.lastname: Murray
authors.forename: H L
authors.initials: HL
authors.lastname: Rubin
authors.forename: W M
authors.initials: WM
authors.lastname: Taylor
authors.forename: J T
authors.initials: JT
authors.lastname: Prchal
authors.forename: S K
authors.initials: SK
authors.lastname: Ballas
authors.forename: L M
authors.initials: LM
authors.lastname: Snyder
authors.forename: L
authors.initials: L
authors.lastname: Chrobak
authors.forename: W D
authors.initials: WD
authors.lastname: Melrose
authors.forename: V
authors.initials: V
authors.lastname: Brabec
authors.forename: J
authors.initials: J
authors.lastname: Palek
mesh_terms: D000483:Alleles; D001457:Anion Exchange Protein 1, Erythrocyte / Q000737:chemistry / Q000172:deficiency / Q000235:genetics*; D017487:Ankyrins / Q000172:deficiency / Q000235:genetics; D004252:DNA Mutational Analysis; D004910:Erythrocyte Membrane / Q000737:chemistry; D016368:Frameshift Mutation; D015870:Gene Expression; D006801:Humans; D009154:Mutation; D010641:Phenotype; D017354:Point Mutation; D016133:Polymerase Chain Reaction; D018807:Polymorphism, Single-Stranded Conformational; D011487:Protein Conformation; D012326:RNA Splicing; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D017384:Sequence Deletion; D013049:Spectrin / Q000172:deficiency / Q000235:genetics; D013103:Spherocytosis, Hereditary / Q000145:classification / Q000235:genetics*
chemical_list: D001457:Anion Exchange Protein 1, Erythrocyte; D017487:Ankyrins; D012333:RNA, Messenger; D013049:Spectrin