Laminin-5 and hemidesmosomes: role of the alpha 3 chain subunit in hemidesmosome stability and assembly.

Hemidesmosomes are complex macromolecular structures which integrate elements of the extracellular matrix and the cytoskeleton of epithelial cells. To characterize cell-matrix interactions in the hemidesmosome, we have made use of 804G cells which possess the unusual ability to assemble hemidesmosomes in vitro. During the course of our studies, we have raised a set of monoclonal antibodies against rat laminin-5, the major structural element comprising 804G matrix. One of these, termed CM6, recognizes the 150 kDa alpha chain of rat laminin-5 and binds the globular (G) domain of intact laminin-5 molecules as determined by rotary shadowing. CM6 antibodies perturb formed hemidesmosomes in 804G cells. In particular, within 1 hour of incubation of 804G cells with CM6 antibodies, colocalization of laminin-5 and alpha 6 beta 4 integrin is lost and by 2 hours, staining generated by hemidesmosomal antibodies appears primarily cytoplasmic in the perinuclear zone. Ultrastructurally, CM6 antibodies first appear to induce detachment of hemidesmosomes from the underlying matrix. Next, portions of the basal cell surface invaginate to form vesicles whose cytoplasmic-facing surface is coated with hemidesmosomes still associated with keratin intermediate filaments. Anchoring filaments extend into the inside compartment of the vesicles. We have also studied the impact of CM6 antibodies on a model system in which the matrix of 804G cells induces de novo assembly of hemidesmosomes in human keratinocytes. This process involves the plasma membrane reorganization of the hemidesmosome associated integrin alpha 6 beta 4 as well as a redistribution of other hemidesmosome components such as the 230 kDa bullous pemphigoid antigen. Pretreatment of 804G matrix with CM6 antibodies blocks such plasma membrane reorganization of hemidesmosome components and inhibits hemidesmosome formation. Our studies indicate a crucial role for the G domain of the alpha chain of laminin-5 in both nucleation of hemidesmosome assembly as well as maintenance of hemidesmosome structural integrity.

[1]  J. Falk-Marzillier,et al.  Rapid spreading and mature hemidesmosome formation in HaCaT keratinocytes induced by incubation with soluble laminin-5r. , 1995, The Journal of investigative dermatology.

[2]  J. Schlessinger,et al.  Signal transduction by the α6β4 integrin: distinct β4 subunit sites mediate recruitment of Shc/Grb2 and association with the cytoskeleton of hemidesmosomes , 1995 .

[3]  A. Aguzzi,et al.  Cloning and Complete Primary Structure of the Mouse Laminin α3 Chain , 1995, The Journal of Biological Chemistry.

[4]  S. Baker,et al.  Molecular genetic studies of a human epidermal autoantigen (the 180-kD bullous pemphigoid antigen/BP180): identification of functionally important sequences within the BP180 molecule and evidence for an interaction between BP180 and alpha 6 integrin , 1995, The Journal of cell biology.

[5]  J. R. McMillan,et al.  A homozygous nonsense mutation in the alpha 3 chain gene of laminin 5 (LAMA3) in lethal (Herlitz) junctional epidermolysis bullosa. , 1995, Human molecular genetics.

[6]  M. Bissell,et al.  Laminin mediates tissue-specific gene expression in mammary epithelia , 1995 .

[7]  E. Fuchs,et al.  Gene targeting of BPAG1: Abnormalities in mechanical strength and cell migration in stratified epithelia and neurologic degeneration , 1995, Cell.

[8]  F. Giancotti,et al.  A recombinant tail-less integrin beta 4 subunit disrupts hemidesmosomes, but does not suppress alpha 6 beta 4-mediated cell adhesion to laminins , 1995, The Journal of cell biology.

[9]  R. Burgeson,et al.  A homozygous nonsense mutation in the beta 3 chain gene of laminin 5 (LAMB3) in Herlitz junctional epidermolysis bullosa. , 1994, Genomics.

[10]  W. Carter,et al.  Cloning of the LamA3 gene encoding the alpha 3 chain of the adhesive ligand epiligrin. Expression in wound repair. , 1994, The Journal of biological chemistry.

[11]  Jennifer Asmuth,et al.  Hemidesmosomes: extracellular matrix/intermediate filament connectors. , 1994, Experimental cell research.

[12]  S. Ferrari,et al.  Expression and function of nerve growth factor and nerve growth factor receptor on cultured keratinocytes. , 1994, The Journal of investigative dermatology.

[13]  J. Sanes,et al.  A new nomenclature for the laminins. , 1994, Matrix biology : journal of the International Society for Matrix Biology.

[14]  M. Aumailley,et al.  Kalinin is more efficient than laminin in promoting adhesion of primary keratinocytes and some other epithelial cells and has a different requirement for integrin receptors , 1994, The Journal of cell biology.

[15]  A. Sonnenberg,et al.  The α6β4 Integrin Is a Receptor for both Laminin and Kalinin , 1994 .

[16]  M. Matter,et al.  A novel laminin E8 cell adhesion site required for lung alveolar formation in vitro , 1994, The Journal of cell biology.

[17]  J. Uitto,et al.  Herlitz's junctional epidermolysis bullosa is linked to mutations in the gene (LAMC2) for the γ2 subunit of nicein/kalinin (LAMININ–5) , 1994, Nature Genetics.

[18]  J. Uitto,et al.  Mutations in the γ2 chain gene (LAMC2) of kalinin/laminin 5 in the junctional forms of epidermolysis bullosa , 1994, Nature Genetics.

[19]  J. Troy,et al.  A passive transfer model of the organ-specific autoimmune disease, bullous pemphigoid, using antibodies generated against the hemidesmosomal antigen, BP180. , 1993, The Journal of clinical investigation.

[20]  K. Tryggvason,et al.  The laminin family. , 1993, Current opinion in cell biology.

[21]  J. C. Jones,et al.  The matrix secreted by 804G cells contains laminin-related components that participate in hemidesmosome assembly in vitro. , 1993, Journal of cell science.

[22]  Y. Yamada,et al.  Recombinant laminin G domain mediates myoblast adhesion and heparin binding. , 1993, The Journal of biological chemistry.

[23]  T. Nishikawa,et al.  Further analyses of epitopes for human monoclonal anti-basement membrane zone antibodies produced by stable human hybridoma cell lines constructed with Epstein-Barr virus transformants. , 1993, The Journal of investigative dermatology.

[24]  R. Eddy,et al.  A truncated laminin chain homologous to the B2 chain: structure, spatial expression, and chromosomal assignment , 1992, The Journal of cell biology.

[25]  R. Burgeson,et al.  The anchoring filament protein kalinin is synthesized and secreted as a high molecular weight precursor. , 1992, The Journal of biological chemistry.

[26]  V. Quaranta,et al.  Surface relocation of alpha 6 beta 4 integrins and assembly of hemidesmosomes in an in vitro model of wound healing , 1991, The Journal of cell biology.

[27]  A. Skubitz,et al.  Synthetic peptides from the carboxy-terminal globular domain of the A chain of laminin: their ability to promote cell adhesion and neurite outgrowth, and interact with heparin and the beta 1 integrin subunit , 1991, The Journal of cell biology.

[28]  R. Burgeson,et al.  Kalinin: an epithelium-specific basement membrane adhesion molecule that is a component of anchoring filaments , 1991, The Journal of cell biology.

[29]  V. Quaranta,et al.  A function for the integrin alpha 6 beta 4 in the hemidesmosome. , 1991, Cell regulation.

[30]  W. Carter,et al.  Epiligrin, a new cell adhesion ligand for integrin α3β1 in epithelial basement membranes , 1991, Cell.

[31]  M Loizidou,et al.  Integrin alpha 6/beta 4 complex is located in hemidesmosomes, suggesting a major role in epidermal cell-basement membrane adhesion , 1991, The Journal of cell biology.

[32]  E. Maizels,et al.  Isozymes of cAMP-dependent protein kinase present in the rat corpus luteum. , 1991, The Journal of biological chemistry.

[33]  J. C. Jones,et al.  Formation of hemidesmosomes in vitro by a transformed rat bladder cell line , 1991, The Journal of cell biology.

[34]  M. Stepp,et al.  Alpha 6 beta 4 integrin heterodimer is a component of hemidesmosomes. , 1990, Proceedings of the National Academy of Sciences of the United States of America.

[35]  V. Quaranta,et al.  Epithelial integrin alpha 6 beta 4: complete primary structure of alpha 6 and variant forms of beta 4 , 1990, The Journal of cell biology.

[36]  E. Engvall,et al.  Merosin, a tissue-specific basement membrane protein, is a laminin-like protein. , 1990, Proceedings of the National Academy of Sciences of the United States of America.

[37]  C. R. Jones,et al.  Immunochemical characterization of three components of the hemidesmosome and their expression in cultured epithelial cells , 1989, The Journal of cell biology.

[38]  J. Ortonne,et al.  Monoclonal antibody GB3, a new probe for the study of human basement membranes and hemidesmosomes. , 1987, Experimental cell research.

[39]  G. Heijne.  A new method for predicting signal sequence cleavage sites. , 1986 .

[40]  F. Sanger,et al.  DNA sequencing with chain-terminating inhibitors. , 1977, Proceedings of the National Academy of Sciences of the United States of America.

[41]  W. Carter,et al.  Rapid Epiligrin, the Major Human Keratinocyte Integrin Ligand, Is a Target in Both an Acquired Autoimmune and an Inherited Subepidermal Blistering Skin Disease , 2022 .