title: BPAG1-e restricts keratinocyte migration through control of adhesion stability.
abstract: Bullous pemphigoid antigen 1 (BPAG1-e, also known as BP230) is a member of the plakin family of hemidesmosome cytoskeletal linker proteins that is encoded by an isoform of the dystonin (DST) gene. Recently, we reported two unrelated families with homozygous nonsense mutations in this DST isoform that led to ultrastructural loss of hemidesmosomal inner plaques and clinical features of trauma-induced skin fragility. We now demonstrate that keratinocytes isolated from these individuals have significant defects in adhesion, as well as increased cell spreading and migration. These mutant keratinocytes also display reduced levels of β4 integrins at the cell surface but increased total protein levels of keratin-14 and β1 integrins. These alterations in cell behavior and protein expression were not seen in control keratinocytes in which BPAG1-e expression had been silenced by stable expression of short hairpin RNA to target DST. The failure of knockdown approaches to recapitulate the changes in morphology, adhesion, and migration seen in patient cells therefore suggests such approaches are not appropriate to study loss of this protein in vivo. The contrasting findings in keratinocytes harboring naturally occurring mutations, however, demonstrate a previously unappreciated key role for BPAG1-e in regulating keratinocyte adhesion and migration and suggest a requirement for this protein in controlling functional switching between integrin types in epithelial cells.
authors.affiliation: Randall Division of Cell and Molecular Biophysics, King's College London, Guy's Campus, London, UK; St Johns Institute of Dermatology, King's College London, Guys Campus, London, UK; Division of Molecular Cell Biology, Institute for Molecular Bioscience, The University of Queensland, St Lucia, Brisbane, Queensland, Australia.
authors.forename: Magdalene
authors.initials: M
authors.lastname: Michael
authors.affiliation: Randall Division of Cell and Molecular Biophysics, King's College London, Guy's Campus, London, UK; St Johns Institute of Dermatology, King's College London, Guys Campus, London, UK.
authors.forename: Rumena
authors.initials: R
authors.lastname: Begum
authors.affiliation: St Johns Institute of Dermatology, King's College London, Guys Campus, London, UK.
authors.forename: Kenneth
authors.initials: K
authors.lastname: Fong
authors.affiliation: Jacqui Wood Cancer Centre, University of Dundee, Ninewells Hospital and Medical School, Dundee, UK.
authors.forename: Celine
authors.initials: C
authors.lastname: Pourreyrone
authors.affiliation: Jacqui Wood Cancer Centre, University of Dundee, Ninewells Hospital and Medical School, Dundee, UK.
authors.forename: Andrew P
authors.initials: AP
authors.lastname: South
authors.affiliation: St Johns Institute of Dermatology, King's College London, Guys Campus, London, UK.
authors.forename: John A
authors.initials: JA
authors.lastname: McGrath
authors.affiliation: Randall Division of Cell and Molecular Biophysics, King's College London, Guy's Campus, London, UK. Electronic address: maddy.parsons@kcl.ac.uk.
authors.forename: Maddy
authors.initials: M
authors.lastname: Parsons
mesh_terms: D001940:Breast / Q000166:cytology; D002352:Carrier Proteins / Q000235:genetics* / Q000378:metabolism*; D002448:Cell Adhesion / Q000502:physiology*; D002460:Cell Line; D002465:Cell Movement / Q000502:physiology*; D003598:Cytoskeletal Proteins / Q000235:genetics* / Q000378:metabolism*; D003896:Desmosomes / Q000502:physiology; D000071821:Dystonin; D005260:Female; D006801:Humans; D019012:Integrin beta1 / Q000378:metabolism; D039663:Integrin beta4 / Q000378:metabolism; D053547:Keratin-14 / Q000378:metabolism; D015603:Keratinocytes / Q000166:cytology* / Q000502:physiology; D008875:Middle Aged; D009154:Mutation; D009419:Nerve Tissue Proteins / Q000235:genetics* / Q000378:metabolism*
chemical_list: D002352:Carrier Proteins; D003598:Cytoskeletal Proteins; C486268:DST protein, human; D000071821:Dystonin; D019012:Integrin beta1; D039663:Integrin beta4; C507540:KRT14 protein, human; D053547:Keratin-14; D009419:Nerve Tissue Proteins
references.citation: Exp Cell Res. 2007 Jun 10;313(10):2189-203
references.pmid: 17499243
references.citation: J Cell Biol. 2001 Aug 20;154(4):691-7
references.pmid: 11514586
references.citation: J Am Acad Dermatol. 2008 Jun;58(6):931-50
references.pmid: 18374450
references.citation: J Dermatol Sci. 2011 Apr;62(1):1-7
references.pmid: 21376539
references.citation: Exp Cell Res. 2008 Sep 10;314(15):2750-61
references.pmid: 18638474
references.citation: J Invest Dermatol. 1999 Apr;112(4):411-8
references.pmid: 10201522
references.citation: Mol Biol Cell. 2009 Jun;20(12):2954-62
references.pmid: 19403692
references.citation: J Am Acad Dermatol. 2009 Sep;61(3):367-84; quiz 385-6
references.pmid: 19700010
references.citation: Exp Cell Res. 2003 Feb 1;283(1):67-79
references.pmid: 12565820
references.citation: J Invest Dermatol. 2013 Sep;133(9):2121-6
references.pmid: 23949764
references.citation: J Cell Sci. 2000 Sep;113 ( Pt 17):3051-62
references.pmid: 10934043
references.citation: J Pathol. 2005 Mar;205(4):468-75
references.pmid: 15685705
references.citation: Cell. 1995 Apr 21;81(2):233-43
references.pmid: 7736575
references.citation: J Invest Dermatol. 2013 Jan;133(1):181-90
references.pmid: 22895363
references.citation: Nat Rev Mol Cell Biol. 2009 Jul;10(7):445-57
references.pmid: 19546857
references.citation: Cell. 1991 Sep 20;66(6):1301-11
references.pmid: 1717157
references.citation: J Cell Sci. 2003 Jan 15;116(Pt 2):387-99
references.pmid: 12482924
references.citation: FASEB J. 2010 Nov;24(11):4133-52
references.pmid: 20624931
references.citation: J Cell Sci. 1995 Nov;108 ( Pt 11):3463-71
references.pmid: 8586658
references.citation: Exp Cell Res. 2006 Sep 10;312(15):2712-25
references.pmid: 16797530
references.citation: J Invest Dermatol. 2010 Jun;130(6):1551-7
references.pmid: 20164846
references.citation: Science. 2003 Dec 5;302(5651):1704-9
references.pmid: 14657486
references.citation: Cell Motil Cytoskeleton. 2007 Dec;64(12):897-905
references.pmid: 17849487
references.citation: J Am Acad Dermatol. 2009 Sep;61(3):387-402; quiz 403-4
references.pmid: 19700011
references.citation: Trends Cell Biol. 2006 Jul;16(7):376-83
references.pmid: 16757171
references.citation: J Cell Sci. 2001 Oct;114(Pt 19):3409-10
references.pmid: 11682600
references.citation: J Invest Dermatol. 2012 Mar;132(3 Pt 1):742-4
references.pmid: 22113475
references.citation: Development. 2002 May;129(9):2303-15
references.pmid: 11959837