title: Phosphorylation of threonine 1736 in the C-terminal tail of integrin β4 contributes to hemidesmosome disassembly.
abstract: During wound healing, hemidesmome disassembly enables keratinocyte migration and proliferation. Hemidesmosome dynamics are altered downstream of epidermal growth factor (EGF) receptor activation, following the phosphorylation of integrin β4 residues S1356 and S1364, which reduces the interaction with plectin; however, this event is insufficient to drive complete hemidesmome disassembly. In the studies reported here, we used a fluorescence resonance energy transfer-based assay to demonstrate that the connecting segment and carboxy-terminal tail of the β4 cytoplasmic domain interact, which facilitates the formation of a binding platform for plectin. In addition, analysis of a β4 mutant containing a phosphomimicking aspartic acid residue at T1736 in the C-tail suggests that phosphorylation of this residue regulates the interaction with the plectin plakin domain. The aspartic acid mutation of β4 T1736 impaired hemidesmosome formation in junctional epidermolysis associated with pyloric atresia/β4 keratinocytes. Furthermore, we show that T1736 is phosphorylated downstream of protein kinase C and EGF receptor activation and is a substrate for protein kinase D1 in vitro and in cells, which requires its translocation to the plasma membrane and subsequent activation. In conclusion, we identify T1736 as a novel phosphorylation site that contributes to the regulation of hemidesmome disassembly, a dynamically regulated process involving the concerted phosphorylation of multiple β4 residues.
authors.affiliation: Division of Cell Biology, The Netherlands Cancer Institute, 1066 CX Amsterdam, The Netherlands.
authors.forename: Evelyne
authors.initials: E
authors.lastname: Frijns
authors.forename: Ingrid
authors.initials: I
authors.lastname: Kuikman
authors.forename: Sandy
authors.initials: S
authors.lastname: Litjens
authors.forename: Marcel
authors.initials: M
authors.lastname: Raspe
authors.forename: Kees
authors.initials: K
authors.lastname: Jalink
authors.forename: Michael
authors.initials: M
authors.lastname: Ports
authors.forename: Kevin
authors.initials: K
authors.lastname: Wilhelmsen
authors.forename: Arnoud
authors.initials: A
authors.lastname: Sonnenberg
mesh_terms: D000818:Animals; D019556:COS Cells; D002460:Cell Line; D002462:Cell Membrane / Q000378:metabolism; D002465:Cell Movement; D049109:Cell Proliferation; D002522:Chlorocebus aethiops; D004815:Epidermal Growth Factor / Q000378:metabolism; D066246:ErbB Receptors / Q000235:genetics; D031541:Fluorescence Resonance Energy Transfer; D057809:HEK293 Cells; D022002:Hemidesmosomes / Q000378:metabolism* / Q000648:ultrastructure; D006801:Humans; D039663:Integrin beta4 / Q000235:genetics / Q000378:metabolism*; D015603:Keratinocytes / Q000378:metabolism*; D009154:Mutation; D010766:Phosphorylation; D051190:Plectin / Q000378:metabolism; D011493:Protein Kinase C / Q000235:genetics / Q000378:metabolism; D013912:Threonine / Q000378:metabolism
chemical_list: D039663:Integrin beta4; D051190:Plectin; D013912:Threonine; D004815:Epidermal Growth Factor; C089212:protein kinase D; D066246:ErbB Receptors; D011493:Protein Kinase C