title: A crucial role for β2 integrins in podosome formation, dynamics and Toll-like-receptor-signaled disassembly in dendritic cells.
abstract: The dynamic properties of podosomes, their ability to degrade the underlying matrix and their modulation by Toll-like receptor (TLR) signaling in dendritic cells (DCs) suggests they have an important role in migration. Integrins are thought to participate in formation and dynamics of podosomes but the multiplicity of integrins in podosomes has made this difficult to assess. We report that murine DCs that lack β2 integrins fail to form podosomes. Re-expression of β2 integrins restored podosomes but not when the membrane proximal or distal NPxF motifs, or when an intervening triplet of threonine residues were mutated. We show that β2 integrins are remarkably long-lived in podosome clusters and form a persistent framework that hosts multiple actin-core-formation events at the same or adjacent sites. When β2 integrin amino acid residues 745 or 756 were mutated from Ser to Ala, podosomes became resistant to dissolution mediated through TLR signaling. TLR signaling did not detectably modulate phosphorylation at these sites but mutation of either residue to phospho-mimetic Asp increased β2 integrin turnover in podosomes, indicating that phosphorylation at one or both sites establishes permissive conditions for TLR-signaled podosome disassembly.
authors.affiliation: Division of Cell Biology and Immunology, College of Life Sciences, University of Dundee, Dundee DD1 5EH, UK.
authors.forename: Christian
authors.initials: C
authors.lastname: Gawden-Bone
authors.affiliation: Division of Cell Biology and Immunology, College of Life Sciences, University of Dundee, Dundee DD1 5EH, UK.
authors.forename: Michele A
authors.initials: MA
authors.lastname: West
authors.affiliation: University of Dundee, Ninewells Hospital and Medical School, Dundee DD1 9SY, UK.
authors.forename: Vicky L
authors.initials: VL
authors.lastname: Morrison
authors.affiliation: Division of Cell Biology and Immunology, College of Life Sciences, University of Dundee, Dundee DD1 5EH, UK.
authors.forename: Alexander J
authors.initials: AJ
authors.lastname: Edgar
authors.affiliation: Division of Cell Biology and Immunology, College of Life Sciences, University of Dundee, Dundee DD1 5EH, UK.
authors.forename: Sarah J
authors.initials: SJ
authors.lastname: McMillan
authors.affiliation: MRC Protein Phosphorylation and Ubiquitylation Unit, College of Life Sciences, University of Dundee, Dundee DD1 5EH, UK.
authors.forename: Brian D
authors.initials: BD
authors.lastname: Dill
authors.affiliation: MRC Protein Phosphorylation and Ubiquitylation Unit, College of Life Sciences, University of Dundee, Dundee DD1 5EH, UK.
authors.forename: Matthias
authors.initials: M
authors.lastname: Trost
authors.affiliation: Division of Cell Biology and Immunology, College of Life Sciences, University of Dundee, Dundee DD1 5EH, UK.
authors.forename: Alan
authors.initials: A
authors.lastname: Prescott
authors.affiliation: University of Dundee, Ninewells Hospital and Medical School, Dundee DD1 9SY, UK.
authors.forename: Susanna C
authors.initials: SC
authors.lastname: Fagerholm
authors.affiliation: Division of Cell Biology and Immunology, College of Life Sciences, University of Dundee, Dundee DD1 5EH, UK c.watts@dundee.ac.uk.
authors.forename: Colin
authors.initials: C
authors.lastname: Watts
mesh_terms: D000818:Animals; D018821:CD18 Antigens / Q000378:metabolism*; D021961:Cell Membrane Structures / Q000378:metabolism; D002465:Cell Movement / Q000502:physiology; D003713:Dendritic Cells / Q000166:cytology* / Q000378:metabolism*; D005260:Female; D051379:Mice; D008810:Mice, Inbred C57BL; D011247:Pregnancy; D015398:Signal Transduction; D051193:Toll-Like Receptors / Q000378:metabolism*
chemical_list: D018821:CD18 Antigens; D051193:Toll-Like Receptors
keywords: Dendritic cells; Integrins; Podosomes
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