Regulation of keratinocyte terminal differentiation by integrin-extracellular matrix interactions.
Suspension-induced terminal differentiation of human epidermal keratinocytes can be inhibited by fibronectin through binding to the alpha 5 beta 1 integrin. We have investigated the effect of fibronectin on expression of integrins and proteins of the actin cytoskeleton and have explored the nature of the differentiation stimulus by testing different combinations of anti-integrin monoclonal antibodies or extracellular matrix proteins in the suspension assay. Fibronectin prolonged cell surface expression of beta 1 integrins but did not overcome the inhibition of intracellular transport of integrins that occurs when keratinocytes are placed in suspension. Fibronectin did not prevent the suspension-induced decline in the level of mRNAs encoding the beta 1 integrin subunit, actin, filamin and alpha-actinin; furthermore, the inhibition of terminal differentiation did not depend on the state of assembly of microfilaments or microtubules. Terminal differentiation could be partially inhibited by an adhesion-blocking monoclonal antibody to the beta 1 integrin subunit or by a combination of adhesion blocking antibodies recognising the alpha subunits that associate with beta 1 (alpha 2, alpha 3 and alpha 5). Although laminin and type IV collagen do not inhibit terminal differentiation individually, they were inhibitory when added to cells in combination with a low concentration of fibronectin. We conclude that the proportion of keratinocyte beta 1 integrins occupied by ligand can regulate the initiation of terminal differentiation independently of the state of assembly of the actin cytoskeleton.
Watt
F M
FM
Keratinocyte Laboratory, Imperial Cancer Research Fund, London, UK.
Kubler
M D
MD
Hotchin
N A
NA
Nicholson
L J
LJ
Adams
J C
JC
D000199:Actins / Q000378:metabolism; D000911:Antibodies, Monoclonal / Q000276:immunology; D002454:Cell Differentiation / Q000187:drug effects; D002478:Cells, Cultured; D003094:Collagen / Q000494:pharmacology; D003599:Cytoskeleton / Q000187:drug effects / Q000648:ultrastructure; D005109:Extracellular Matrix / Q000502:physiology*; D016326:Extracellular Matrix Proteins / Q000494:pharmacology; D005353:Fibronectins / Q000378:metabolism / Q000494:pharmacology*; D006801:Humans; D039201:Integrin alpha3beta1; D016023:Integrins / Q000276:immunology / Q000378:metabolism / Q000502:physiology*; D015603:Keratinocytes / Q000166:cytology* / Q000187:drug effects; D007797:Laminin / Q000494:pharmacology; D011485:Protein Binding; D011498:Protein Precursors / Q000096:biosynthesis; D012333:RNA, Messenger / Q000096:biosynthesis; D017469:Receptors, Fibronectin
D000199:Actins; D000911:Antibodies, Monoclonal; D016326:Extracellular Matrix Proteins; D005353:Fibronectins; D039201:Integrin alpha3beta1; D016023:Integrins; D007797:Laminin; D011498:Protein Precursors; D012333:RNA, Messenger; D017469:Receptors, Fibronectin; C032253:involucrin; D003094:Collagen