title: Phosphorylation within the cysteine-rich region of dystrophin enhances its association with β-dystroglycan and identifies a potential novel therapeutic target for skeletal muscle wasting.
abstract: Mutations in dystrophin lead to Duchenne muscular dystrophy, which is among the most common human genetic disorders. Dystrophin nucleates assembly of the dystrophin-glycoprotein complex (DGC), and a defective DGC disrupts an essential link between the intracellular cytoskeleton and the basal lamina, leading to progressive muscle wasting. In vitro studies have suggested that dystrophin phosphorylation may affect interactions with actin or syntrophin, yet whether this occurs in vivo or affects protein function remains unknown. Utilizing nanoflow liquid chromatography mass spectrometry, we identified 18 phosphorylated residues within endogenous dystrophin. Mutagenesis revealed that phosphorylation at S3059 enhances the dystrophin-dystroglycan interaction and 3D modeling utilizing the Rosetta software program provided a structural model for how phosphorylation enhances this interaction. These findings demonstrate that phosphorylation is a key mechanism regulating the interaction between dystrophin and the DGC and reveal that posttranslational modification of a single amino acid directly modulates the function of dystrophin.
authors.affiliation: Basic and Clinical Myology Laboratory, Department of Physiology, University of Melbourne, VIC 3010, Australia Department of Neurology, University of Washington School of Medicine, Seattle, WA 98195-7720, USA.
authors.forename: Kristy
authors.initials: K
authors.lastname: Swiderski
authors.affiliation: Department of Medicinal Chemistry, University of Washington School of Medicine, Seattle, WA 98195-7610, USA.
authors.forename: Scott A
authors.initials: SA
authors.lastname: Shaffer
authors.affiliation: Department of Medicinal Chemistry, University of Washington School of Medicine, Seattle, WA 98195-7610, USA.
authors.forename: Byron
authors.initials: B
authors.lastname: Gallis
authors.affiliation: Department of Neurology, University of Washington School of Medicine, Seattle, WA 98195-7720, USA.
authors.forename: Guy L
authors.initials: GL
authors.lastname: Odom
authors.affiliation: Department of Neurology, University of Washington School of Medicine, Seattle, WA 98195-7720, USA.
authors.forename: Andrea L
authors.initials: AL
authors.lastname: Arnett
authors.affiliation: Department of Medicinal Chemistry, University of Washington School of Medicine, Seattle, WA 98195-7610, USA.
authors.forename: J
authors.initials: J
authors.lastname: Scott Edgar
authors.affiliation: Basic and Clinical Myology Laboratory, Department of Physiology, University of Melbourne, VIC 3010, Australia.
authors.forename: Dale M
authors.initials: DM
authors.lastname: Baum
authors.affiliation: Basic and Clinical Myology Laboratory, Department of Physiology, University of Melbourne, VIC 3010, Australia.
authors.forename: Annabel
authors.initials: A
authors.lastname: Chee
authors.affiliation: Basic and Clinical Myology Laboratory, Department of Physiology, University of Melbourne, VIC 3010, Australia.
authors.forename: Timur
authors.initials: T
authors.lastname: Naim
authors.affiliation: Muscle Biology and Therapeutics Laboratory, Baker IDI Heart and Diabetes Institute, Melbourne, VIC 3004, Australia.
authors.forename: Paul
authors.initials: P
authors.lastname: Gregorevic
authors.affiliation: Basic and Clinical Myology Laboratory, Department of Physiology, University of Melbourne, VIC 3010, Australia.
authors.forename: Kate T
authors.initials: KT
authors.lastname: Murphy
authors.affiliation: Department of Biochemistry, University of Washington School of Medicine, Seattle, WA 98195-7350, USA and Program in Molecular and Cellular Biology, University of Washington School of Medicine, Seattle, WA 98195-7275, USA.
authors.forename: James
authors.initials: J
authors.lastname: Moody
authors.affiliation: Department of Medicinal Chemistry, University of Washington School of Medicine, Seattle, WA 98195-7610, USA.
authors.forename: David R
authors.initials: DR
authors.lastname: Goodlett
authors.affiliation: Basic and Clinical Myology Laboratory, Department of Physiology, University of Melbourne, VIC 3010, Australia.
authors.forename: Gordon S
authors.initials: GS
authors.lastname: Lynch
authors.affiliation: Department of Neurology, University of Washington School of Medicine, Seattle, WA 98195-7720, USA Department of Biochemistry, University of Washington School of Medicine, Seattle, WA 98195-7350, USA and Program in Molecular and Cellular Biology, University of Washington School of Medicine, Seattle, WA 98195-7275, USA jsc5@uw.edu.
authors.forename: Jeffrey S
authors.initials: JS
authors.lastname: Chamberlain
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D002454:Cell Differentiation; D002460:Cell Line; D003545:Cysteine / Q000737:chemistry / Q000378:metabolism; D049030:Dystroglycans / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D016189:Dystrophin / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D049029:Dystrophin-Associated Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D005786:Gene Expression Regulation; D057809:HEK293 Cells; D006801:Humans; D051379:Mice; D008810:Mice, Inbred C57BL; D018101:Mice, Inbred mdx; D008958:Models, Molecular; D008969:Molecular Sequence Data; D018482:Muscle, Skeletal / Q000378:metabolism* / Q000473:pathology; D009133:Muscular Atrophy / Q000235:genetics / Q000378:metabolism* / Q000473:pathology; D032446:Myoblasts / Q000166:cytology / Q000378:metabolism; D010766:Phosphorylation; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D012694:Serine / Q000737:chemistry / Q000378:metabolism; D015398:Signal Transduction
chemical_list: D016189:Dystrophin; D049029:Dystrophin-Associated Proteins; D011993:Recombinant Fusion Proteins; C091824:syntrophin; D049030:Dystroglycans; D012694:Serine; D003545:Cysteine
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