Low frequency of ankyrin mutations in hereditary spherocytosis: Identification of three novel mutations

Leite, Roberta Campanile A.; Bassères, Daniela S.; Ferreira, Jovino S.; Alberto, Fernando; Costa, Fernando Ferreira; Saad, Sara T.O.

doi:10.1002/1098-1004(200012)16:6<529::aid-humu13>3.0.co;2-n

Cited by 17 publications

(17 citation statements)

References 12 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Similarly, the genetic bases of HS in non-European populations have not been defined. The exceptions are Japan and Brazil, where mutation screening has been performed [35,36]. These studies found that ankyrin gene mutations were less common than in Europeans.…”

Section: Discussionmentioning

confidence: 99%

Ankyrin‐linked hereditary spherocytosis in an African–American kindred

et al. 2008

View full text Add to dashboard Cite

Mutations of ankyrin-1 are the most frequent cause of the inherited hemolytic anemia, hereditary spherocytosis (HS), in people of European ancestry. Ankyrin-1, which provides the primary linkage between the erythrocyte membrane skeleton and the plasma membrane, has numerous isoforms generated by alternative splicing, alternate polyadenylation, use of tissue-specific promoters, and alternate NH 2 or COOH-termini. Mutation detection in erythrocyte membrane protein genes, including ankyrin, has been a challenge, primarily due to the large size of these genes, and the apparent frequent occurrence of HS-associated null alleles. Using denaturing high-performance liquid chromatography (DHPLC), we screened the ankyrin gene of the proband of a large, three generation African-American kindred with ankyrin-deficient HS. DHPLC yielded an abnormal chromatogram for exon 1. Examination of the corresponding exon 1 sequence in genomic DNA from the proband revealed heterozygosity for a mutation of the initiator methionine (ATG to ATA Met 1 Ile). Coupled in vitro transcription/translation studies with rabbit reticulocyte lysates demonstrated that the wild-type ankyrin erythroid cDNA initiates only from the known initiator methionine, indicating that the use of alternate initiator methionine is not a mechanism of isoform diversity in erythroid cells. The mutant ankyrin allele, unlike some initiator methionine mutations that utilize downstream codons for translation initiation, was associated with a null allele. This is the first report describing ankyrin-linked HS in an African-American kindred. Am. J. Hematol. 83:789-794, 2008. V

show abstract

Section: Discussionmentioning

confidence: 99%

Ankyrin‐linked hereditary spherocytosis in an African–American kindred

et al. 2008

View full text Add to dashboard Cite

show abstract

“…4) even when yeasts were grown in less stringent medium lacking histidine, leucine, and tryptophane but containing adenine (data not shown). A survey of the literature revealed that one missense mutation (H276R) in the D2 domain is associated with the HS phenotype of one Brazilian patient (35). However, as analyzed by twohybrid assay, this mutation has no effect on Rh/RhAG-AnkR interaction (not shown).…”

Section: The C-terminal Cytoplasmic Domains Of Rh and Rhag Bind To Thmentioning

confidence: 98%

“…Potential interaction between CD47 and protein 4.2 is suggested from the deficiency of CD47 in 4.2(Ϫ)HS red cells (21,22). In the proposed model, the Rh complex and band 3 are associated with each other and the spectrin-based skeleton through their interaction with ankyrin-R. Bottom, in vitro translated, 35 S-labeled Ank-D2 was incubated with the GST fusion proteins and pulled-down with glutathione-Sepharose 4B beads, as described under "Material and Methods." After washing, the Ank-D2 proteins that remained bound were eluted with glutathione, separated by SDS-PAGE and detected by autoradiography.…”

Section: Fig 4 Yeast Two-hybrid Analysis Of the Interaction Betweenmentioning

confidence: 99%

Rh-RhAG/Ankyrin-R, a New Interaction Site between the Membrane Bilayer and the Red Cell Skeleton, Is Impaired by Rhnull-associated Mutation

Nicolas

Kim

Gane

et al. 2003

Journal of Biological Chemistry

117

109

View full text Add to dashboard Cite

Several studies suggest that the Rh complex represents a major interaction site between the membrane lipid bilayer and the red cell skeleton, but little is known about the molecular basis of this interaction. We report here that ankyrin-R is capable of interacting directly with the C-terminal cytoplasmic domain of Rh and RhAG polypeptides. We first show that the primary defect of ankyrin-R in normoblastosis (nb/nb) spherocytosis mutant mice is associated with a sharp reduction of RhAG and Rh polypeptides. Secondly, our flow cytometric analysis of the Triton X-100 extractability of recombinant fusion proteins expressed in erythroleukemic cell lines suggests that the C-terminal cytoplasmic domains of Rh and RhAG are sufficient to mediate interaction with the erythroid membrane skeleton. Using the yeast two-hybrid system, we demonstrate a direct interaction between the cytoplasmic tails of Rh and RhAG and the second repeat domain (D2) of ankyrin-R. This finding is supported by the demonstration that the substitution of Asp-399 in the cytoplasmic tail of RhAG, a mutation associated with the deficiency of the Rh complex in one Rh null patient, totally impaired interaction with domain D2 of ankyrin-R. These results identify the Rh/RhAG-ankyrin complex as a new interaction site between the red cell membrane and the spectrin-based skeleton, the disruption of which might result in the stomato-spherocytosis typical of Rh null red cells.

show abstract

“…Two linked variants, -108 T→C and -153 G→A, were discovered in several kindreds with ankyrin-deficient spherocytosis who lacked other causative mutations (5)(6)(7). Neither variant is located in known critical regions of promoter function, e.g., transcription initiation sites or transcription factor-binding sites (8).…”

Section: Introductionmentioning

confidence: 99%