title: Gene disruption of dematin causes precipitous loss of erythrocyte membrane stability and severe hemolytic anemia.
abstract: Dematin is a relatively low abundance actin binding and bundling protein associated with the spectrin-actin junctions of mature erythrocytes. Primary structure of dematin includes a loosely folded core domain and a compact headpiece domain that was originally identified in villin. Dematin's actin binding properties are regulated by phosphorylation of its headpiece domain by cyclic adenosine monophosphate-dependent protein kinase. Here, we used a novel gene disruption strategy to generate the whole body dematin gene knockout mouse model (FLKO). FLKO mice, while born at a normal Mendelian ratio, developed severe anemia and exhibited profound aberrations of erythrocyte morphology and membrane stability. Having no apparent effect on primitive erythropoiesis, FLKO mice show significant enhancement of erythroblast enucleation during definitive erythropoiesis. Using membrane protein analysis, domain mapping, electron microscopy, and dynamic deformability measurements, we investigated the mechanism of membrane instability in FLKO erythrocytes. Although many membrane and cytoskeletal proteins remained at their normal levels, the major peripheral membrane proteins spectrin, adducin, and actin were greatly reduced in FLKO erythrocytes. Our results demonstrate that dematin plays a critical role in maintaining the fundamental properties of the membrane cytoskeleton complex.
authors.affiliation: Department of Developmental, Molecular, and Chemical Biology, Tufts University School of Medicine, Boston, MA;
authors.forename: Yunzhe
authors.identifier: 0000-0001-7542-6848
authors.initials: Y
authors.lastname: Lu
authors.affiliation: Department of Developmental, Molecular, and Chemical Biology, Tufts University School of Medicine, Boston, MA;
authors.forename: Toshihiko
authors.initials: T
authors.lastname: Hanada
authors.affiliation: Department of Pediatric Oncology, Dana-Farber Cancer Institute and Division of Hematology/Oncology, Boston Children's Hospital, Harvard Stem Cell Institute, Harvard Medical School, Boston, MA;
authors.forename: Yuko
authors.initials: Y
authors.lastname: Fujiwara
authors.affiliation: Department of Developmental, Molecular, and Chemical Biology, Tufts University School of Medicine, Boston, MA; Pharmacology & Experimental Therapeutics, Tufts University School of Medicine, Boston, MA;
authors.forename: Jennifer O
authors.initials: JO
authors.lastname: Nwankwo
authors.affiliation: Department of Developmental, Molecular, and Chemical Biology, Tufts University School of Medicine, Boston, MA;
authors.forename: Adam J
authors.initials: AJ
authors.lastname: Wieschhaus
authors.affiliation: Division of Hematology, Department of Medicine, Brigham and Women's Hospital, Harvard Medical School, Boston, MA;
authors.forename: John
authors.initials: J
authors.lastname: Hartwig
authors.affiliation: Department of Electrical Engineering and Computer Science and.
authors.forename: Sha
authors.initials: S
authors.lastname: Huang
authors.affiliation: Department of Electrical Engineering and Computer Science and Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, MA; and.
authors.forename: Jongyoon
authors.initials: J
authors.lastname: Han
authors.affiliation: Department of Developmental, Molecular, and Chemical Biology, Tufts University School of Medicine, Boston, MA; Pharmacology & Experimental Therapeutics, Tufts University School of Medicine, Boston, MA; Sackler School of Graduate Biomedical Sciences, Programs in Cellular and Molecular Physiology, and Department of Molecular Microbiology, Tufts University School of Medicine, Boston, MA.
authors.forename: Athar H
authors.identifier: 0000-0003-0335-1861
authors.initials: AH
authors.lastname: Chishti
mesh_terms: D000743:Anemia, Hemolytic / Q000235:genetics / Q000378:metabolism / Q000473:pathology; D000818:Animals; D002148:Calmodulin-Binding Proteins / Q000235:genetics / Q000378:metabolism; D003598:Cytoskeletal Proteins / Q000235:genetics* / Q000378:metabolism; D003599:Cytoskeleton / Q000235:genetics / Q000378:metabolism / Q000473:pathology; D004910:Erythrocyte Membrane / Q000235:genetics / Q000378:metabolism / Q000473:pathology; D005260:Female; D017353:Gene Deletion; D008297:Male; D051379:Mice; D018345:Mice, Knockout; D013049:Spectrin / Q000235:genetics / Q000378:metabolism; D000099291:Adducins
chemical_list: D002148:Calmodulin-Binding Proteins; D003598:Cytoskeletal Proteins; D013049:Spectrin; D000099291:Adducins; C489041:Dmtn protein, mouse