Defective anion transport and marked spherocytosis with membrane instability caused by hereditary total deficiency of red cell band 3 in cattle due to a nonsense mutation

J Clin Invest. 1996 Apr 15;97(8):1804-17. doi: 10.1172/JCI118610.


We studied bovine subjects that exhibited a moderate uncompensated anemia with hereditary spherocytosis inherited in an autosomal incompletely dominant mode and retarded growth. Based on the results of SDS-PAGE, immunoblotting, and electron microscopic analysis by the freeze fracture method, we show here that the proband red cells lacked the band 3 protein completely. Sequence analysis of the proband band 3 cDNA and genomic DNA showed a C --> T substitution resulting in a nonsense mutation (CGA --> TGA; Arg --> Stop) at the position corresponding to codon 646 in human red cell band 3 cDNA. The proband red cells were deficient in spectrin, ankyrin, actin, and protein 4.2, resulting in a distorted and disrupted membrane skeletal network with decreased density. Therefore, the proband red cell membranes were extremely unstable and showed the loss of surface area in several distinct ways such as invagination, vesiculation, and extrusion of microvesicles, leading to the formation of spherocytes. Total deficiency of band 3 also resulted in defective Cl-/HCO3- exchange, causing mild acidosis with decreases in the HCO3- concentration and total CO2 in the proband blood. Our results demonstrate that band 3 indeed contributes to red cell membrane stability, CO2 transport, and acid-base homeostasis, but is not always essential to the survival of this mammal.

Publication types

  • Case Reports
  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Anion Exchange Protein 1, Erythrocyte / deficiency*
  • Anion Exchange Protein 1, Erythrocyte / genetics*
  • Anion Exchange Protein 1, Erythrocyte / isolation & purification
  • Base Sequence
  • Bone Marrow / pathology
  • Cattle
  • Cattle Diseases*
  • Chlorides / blood*
  • Codon
  • DNA Primers
  • DNA, Complementary
  • Electrophoresis, Polyacrylamide Gel
  • Erythrocyte Count
  • Erythrocytes / ultrastructure
  • Female
  • Genes, Dominant
  • Humans
  • Kidney / pathology
  • Microscopy, Electron, Scanning
  • Molecular Sequence Data
  • Point Mutation*
  • Polymerase Chain Reaction
  • Reference Values
  • Spherocytosis, Hereditary / blood
  • Spherocytosis, Hereditary / genetics
  • Spherocytosis, Hereditary / veterinary*


  • Anion Exchange Protein 1, Erythrocyte
  • Chlorides
  • Codon
  • DNA Primers
  • DNA, Complementary