Hereditary spherocytosis with band 3 deficiency. Association with a nonsense mutation of the band 3 gene (allele Lyon), and aggravation by a low-expression allele occurring in trans (allele Genas)

Alloisio, Nicole; Maillet, Philippe; Carré, Geneviève; Texier, Pascale; Vallier, Agnès; Baklouti, Faouzi; Philippe, N; Delaunay, Jean

doi:10.1182/blood.v88.3.1062.1062

Cited by 33 publications

(20 citation statements)

References 21 publications

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“…Approximately 20% of HS cases are due to mutations in AE1. The mutations cause a decrease (20-40%) in total red cell AE1 content due to absence of the mutant protein and an associated decrease in band 4.2 content (3)(4)(5)(6)(7)(8)(9)(10). Studies have been unable to detect either the Prague (10) or the Prague II (5) mutant proteins in mature red cells suggesting that they are unstable and degraded during erythroid development.…”

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Trafficking and Folding Defects in Hereditary Spherocytosis Mutants of the Human Red Cell Anion Exchanger

Quilty¹,

Reithmeier²

2000

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Hereditary spherocytosis (HS) is a common inherited hemolytic anemia caused by mutations in erythrocyte proteins including the anion exchanger, AE1 (band 3). This study examined seven missense mutations (L707P, R760Q, R760W, R808C, H834P, T837M, and R870W) located in the membrane domain of the human AE1 that are associated with this disease. The HS mutants, constructed in full-length AE1 cDNA, could be transiently expressed to similar levels in HEK 293 cells. Immunofluorescence, cell surface biotinylation, and pulse chase labeling showed that the HS mutants all exhibited defective cellular trafficking from the endoplasmic reticulum to the plasma membrane. Impaired binding to an inhibitor affinity matrix indicated that the mutant proteins had non-native structures and may be misfolded. Further characterization of the HS R760Q mutant showed no change in its oligomeric structure or turnover (half-life = 15 h) compared to wild-type AE1, suggesting the mutant was not aggregated or targeted for rapid degradation via the proteasome. Intracellular retention of HS mutant AE1 would lead to destruction of the protein during erythroid development and would account for the lack of HS mutant AE1 in the plasma membrane of the mature red cell.

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Trafficking and Folding Defects in Hereditary Spherocytosis Mutants of the Human Red Cell Anion Exchanger

Quilty¹,

Reithmeier²

2000

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“…Several mutations associated with band 3 deficient HS have been found in the band 3 gene. Amino acid substitutions at conserved positions have been identified (Alloisio et al, 1993;Jarolim et al, 1995Jarolim et al, , 1996Maillet et al, 1995;Eber et al, 1996) but nonsense and frameshift mutations appear to be the most common, and can lead to mRNA instability (Miraglia del Giudice et al, 1997;Jenkins et al, 1996;Alloisio et al, 1996;Jarolim et al, 1996) or abnormal band 3 which is not recovered into the membrane ( Jarolim et al, 1994;Bianchi et al, 1996).…”

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Heterogenous band 3 deficiency in hereditary spherocytosis related to different band 3 gene defects

et al. 1997

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Summary. Among 80 hereditary spherocytosis (HS) kindreds studied using denaturing electrophoretic separation of solubilized eythrocyte membrane proteins, we recognized three prominent subsets: HS with isolated spectrin deficiency, HS with combined spectrin and ankyrin deficiency, and HS with band 3 deficiency. These three subsets represent more than 80% of the HS kindreds studied. In this study, eight dominant HS kindreds with band 3 deficiency were investigated for band 3 mutations. In three of these kindreds, linkage analyses confirmed the band 3 gene as the culprit gene. In an attempt to identify the responsible mutations, denaturing gradient gel electrophoresis (DGGE) was used to explore the coding exons (exons 2-20) of band 3 gene. Five different mutations were found in the eight kindreds. In five kindreds we identified substitutions of highly conserved residues, positioned at boundaries of putative transmembrane segments: a C → T substitution at codon 490 changed arginine (CGC) to cysteine (TGC) in three kindreds, a C → T substitution at codon 837 changed threonine (ACG) to methionine (ATG) in two kindreds. In the sixth kindred a G deletion was found in a stretch of five G starting at position 1475, leading to a stop codon either at position 1527 or 1565. In the seventh kindred a T deletion at position 1600 resulted in a stop codon at position 1733 and in the last kindred a T deletion was identified at position 355, leading to a stop codon at position 447. The mutant transcript was present in HS patients bearing missense mutations, whereas only the normal transcript was found in HS patients with frameshift mutations. In the latter group the mean decrease in membrane band 3 content was significantly lower, leading to speculation that missense mutations may have some sort of dominant negative effect.

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“…Moreover, other variants, with premature stop codons, cannot bind to the membrane either (Jarolim et al, 1994a;Jenkins et al, 1996;Alloisio et al, 1996;Eber et al, 1996). Band 3 Milano, similar to the referred variants (Jarolim et al, 1994b;Jenkins et al, 1996;Alloisio et al, 1996;Eber et al, 1996), is associated with compensated haemolysis, further confirming that the absence of one set of band 3 yields a very mild clinical pattern.…”

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confidence: 63%

“…In most cases the reduction of band 3 amounts to 20-40% and is associated with a secondary decrease in protein 4.2. Several band 3 mutations have recently been detected in HS (Alloisio et al, 1993(Alloisio et al, , 1996Jarolim et al, 1994aJarolim et al, , b, 1995Maillet et al, 1995;Miraglia del Giudice et al, 1995;Eber et al, 1996;Jenkins et al, 1996). We describe a large HS family in which an in-frame insertion of 69 nt, involving the 3 0 end of intron 12 and the 5 0 region of exon 13 was found.…”

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A variant of the EPB3 gene of the anti‐Lepore type in hereditary spherocytosis

et al. 1997

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Summary.The EPB3 gene encodes band 3 (anion exchanger 1) of the red cell membrane. A subset of hereditary spherocytosis (HS) is associated with EPB3 gene mutations and band 3 deficiency. We report a large Italian family in which 10 of the 27 members investigated displayed an autosomal dominant HS. SDS-PAGE revealed a reduction in band 3 in the patients. Screening of the Pst I polymorphic site confirmed the linkage of HS with the EPB3 gene. Analysis of complementary and genomic DNA showed a large additional segment. Nucleotide sequencing disclosed an in-frame duplication of 69 nucleotides (nt) including a triplet of intronic origin and a genuine exonic duplication of 66 nt. Two CCTGC sequences occurred close to one another, one near the intron 12 acceptor splice site (nt ¹7 to ¹3), and the other within exon 13 (nt 1494-1498). We assumed that the abnormal allele arose from an unequal recombination event of the anti-Lepore type between the two CCTGC sequences.At the level of the mutated protein, termed band 3 Milano, the additional segment (Gln plus duplication of residues 478-499) corresponded to the last part of the third transmembrane domain (TM3), the entire second outer loop and part of TM4 as it is currently defined in hydropathy analysis. After deglycosylation of band 3, only the normal band was detected, supporting the view that band 3 Milano is probably not incorporated into the membrane.Keywords: erythrocyte membrane, band 3 protein, hereditary spherocytosis, congenital haemolytic anaemia, DNA duplication.The gene (EPB3) encoding red cell band 3 (anion exchanger 1) maps to 17q21-qter and contains 20 exons (Sahr et al, 1994;Schofield et al, 1994). The size of band 3 mRNA is 4 . 9 kb.Band 3 is the most abundant protein (1 200 000 monomers per cell) of the erythrocyte membrane and plays a key role in maintaining the membrane integrity (Tanner, 1993). The N-terminal cytoplasmic domain (403 amino acids) (Tanner et al, 1988;Lux et al, 1989) anchors the cytoskeleton to the membrane integral domain through interactions with ankyrin and protein 4.2. The transmembrane domain consists of 14 membrane-spanning segments (TM 1-14) connected by inner and outer loops (IL and OL respectively) (Lux et al, 1989).Hereditary spherocytosis (HS), a common haemolytic disorder characterized by a wide heterogeneity at the clinical, biochemical and genetic level , has been related to abnormalities of band 3 as well as those of other red cell membrane proteins such as ankyrin, spectrin and protein 4.2 (Lux & Palek, 1995). HS due to band 3 deficiency has an autosomal dominant mode of transmission and is characterized by mild to moderate anaemia. In most cases the reduction of band 3 amounts to 20-40% and is associated with a secondary decrease in protein 4.2. Several band 3 mutations have recently been detected in HS (Alloisio et al, 1993(Alloisio et al, , 1996Jarolim et al, 1994a Jarolim et al, , b, 1995Maillet et al, 1995;Miraglia del Giudice et al, 1995;Eber et al, 1996;Jenkins et al, 1996). We describe a large HS family in which ...

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Hereditary spherocytosis with band 3 deficiency. Association with a nonsense mutation of the band 3 gene (allele Lyon), and aggravation by a low-expression allele occurring in trans (allele Genas)

Cited by 33 publications

References 21 publications

Trafficking and Folding Defects in Hereditary Spherocytosis Mutants of the Human Red Cell Anion Exchanger

Trafficking and Folding Defects in Hereditary Spherocytosis Mutants of the Human Red Cell Anion Exchanger

Heterogenous band 3 deficiency in hereditary spherocytosis related to different band 3 gene defects

A variant of the EPB3 gene of the anti‐Lepore type in hereditary spherocytosis

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