The cytoskeletal binding domain of band 3 is required for multiprotein complex formation and retention during erythropoiesis

Satchwell, Timothy J.; Hawley, Bethan R.; Bell, Amanda J.; Leticia Ribeiro, M.; Toye, Ashley M.

Abstract

Band 3 is the most abundant protein in the erythrocyte membrane and forms the core of a major multiprotein complex. The absence of band 3 in human erythrocytes has only been reported once, in the homozygous band 3 Coimbra patient. We used in vitro culture of erythroblasts derived from this patient, and separately short hairpin RNA-mediated depletion of band 3, to investigate the development of a band 3-deficient erythrocyte membrane and to specifically assess the stability and retention of band 3 dependent proteins in the absence of this core protein during terminal erythroid differentiation. Further, using lentiviral transduction of N-terminally green fluorescent protein-tagged band 3, we demonstrated the ability to restore expression of band 3 to normal levels and to rescue secondary deficiencies of key proteins including glycophorin A, protein 4.2, CD47 and Rh proteins arising from the absence of band 3 in this patient. By transducing band 3-deficient erythroblasts from this patient with band 3 mutants with absent or impaired ability to associate with the cytoskeleton we also demonstrated the importance of cytoskeletal connectivity for retention both of band 3 and of its associated dependent proteins within the reticulocyte membrane during the process of erythroblast enucleation.

• Journal Article Type: Article
• Acceptance Date: Oct 21, 2014
• Online Publication Date: Jan 1, 2015
• Publication Date: Jan 1, 2015
• Deposit Date: Jul 11, 2024
• Publicly Available Date: Jul 12, 2024
• Journal: Haematologica
• Print ISSN: 0390-6078
• Electronic ISSN: 1592-8721
• Publisher: Ferrata Storti Foundation
• Peer Reviewed: Peer Reviewed
• Volume: 100
• Issue: 1
• Pages: 133-142
• DOI: https://doi.org/10.3324/haematol.2014.114538
• Public URL: https://uwe-repository.worktribe.com/output/12121540