Simultaneous (AC)n microsatellite polymorphism analysis and single-stranded conformation polymorphism screening is an efficient strategy for detecting ankyrin-1 mutations in dominant hereditary spherocytosis

Br J Haematol. 2003 Aug;122(4):669-77. doi: 10.1046/j.1365-2141.2003.04479.x.

Abstract

Nonsense/stop mutations in the ankyrin-1 gene (ANK1) are a major cause of dominant HS (dHS) (frequency of 23% in German dHS patients). To date, no common mutation has been found and therefore a simple mutation screening is not feasible. The reduced expression of one cDNA allele in the (AC)n microsatellite polymorphism of the ankyrin-1 gene, as seen in about 20% of Czech patients with dHS, may identify candidates with a possible frameshift/nonsense mutation. In order to verify the efficiency of this screening we screened the ankyrin-1 gene of 22 Czech dHS patients for both the reduced cDNA allele expression in the frequent (AC)n and the common exonic 26/39 polymorphisms, as well as for polymerase chain reaction (PCR) single-stranded conformation polymorphisms in any one of the 42 exons of ANK1. Anomalous PCR products were sequenced. We found seven new ANK1 frameshift/nonsense mutations in nine patients with, but in none of six patients without, a reduced cDNA allele expression (efficiency of 78%). We conclude that screening of dHS patients for such a reduced allele expression in common ANK1 polymorphisms is an efficient procedure for the identification of candidates for frameshift/nonsense mutations in the ankyrin-1 gene.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Ankyrins / genetics*
  • Codon, Nonsense*
  • DNA Mutational Analysis / methods
  • DNA, Complementary / genetics
  • Erythrocyte Membrane / metabolism
  • Female
  • Frameshift Mutation*
  • Gene Expression
  • Humans
  • Male
  • Microsatellite Repeats / genetics
  • Polymerase Chain Reaction / methods
  • Polymorphism, Genetic
  • Polymorphism, Single-Stranded Conformational
  • RNA, Messenger / genetics
  • Spherocytosis, Hereditary / blood
  • Spherocytosis, Hereditary / genetics*

Substances

  • ANK1 protein, human
  • Ankyrins
  • Codon, Nonsense
  • DNA, Complementary
  • RNA, Messenger