Polarized integrin mediates human keratinocyte adhesion to basal lamina.

Luca, Michele De; Tamura, Ryuichi; Kajiji, Shama; Bondanza, Sergio; Rossino, P; Cancedda, Ranieri; Marchisio, P. C.; Quaranta, Vito

doi:10.1073/pnas.87.17.6888

Cited by 198 publications

(130 citation statements)

References 35 publications

Supporting

Mentioning

125

Contrasting

Order By: Relevance

“…Such an interaction may contribute to the maintenance of cell surface polarity and to the stability of the epithelial sheet. In the stratified squamous epithelia, the receptor is expressed exclusively by the cells of the basal layer (2,4,12,24). Thus, expression of c~, is rapidly turned off at the onset of cell differentiation and movement to the upper cell layers (7).…”

Section: Discussionmentioning

confidence: 99%

“…A similar staining pattern was obtained with the anti-cx~ mAb GOH3, and with the B, mAb 346-11A on mouse skin sections (not shown; see also refs. 4,23,24). The COOH-terminal peptide antibodies generated a different pattern.…”

Section: Immunohistochemical Detection Of B~ and Its Proteolytic Fragmentioning

confidence: 99%

“…T i HE Ot~4 integrin is expressed by epithelial and neuroepithelial cells (22). In epithelial tissues, this integrin is restricted to the basal portion of those cells which contact the basement membrane, suggesting a role in adhesion to basement membranes (4,12,22,24,25). Its ligand may be laminin or a laminin isoform (15, 23).…”

mentioning

confidence: 99%

See 2 more Smart Citations

Proteolytic processing of endogenous and recombinant beta 4 integrin subunit.

Giancotti

Stepp

Suzuki

et al. 1992

The Journal of Cell Biology

122

View full text Add to dashboard Cite

Abstract. The o~4 integrin is a receptor involved in the interaction of epithelial cells with basement membranes. This integrin is unique among the known integrins in that its t4 subunit has a large cytoplasmic domain, The function of this cytoplasmic domain is not known. In this paper we show that the/L subunit undergoes proteolytic processing in cultured cells and provide evidence that this also happens in tissues. Immunoprecipitation experiments indicated that the cytoplasmic domain of #4 is susceptible to a calciumdependent protease present in cellular extracts. In vitro assays with purified calpain showed that this enzyme can cleave ~4 at two distinct sites in the cytoplasmic domain, generating truncated molecules of 165 and 130 kD. Immunoblotting experirnents performed on cultured epithelial cells using an antibody to a peptide modeled after the COOH-terminus of the /L subunit showed 70-kD fragments and several fragments of molecular masses between 185 and 115 kD.Similar fragments were detected in CHO cells transfected with the full-length/L eDNA, but not in control transfected cells or in cells transfected with a mutant eDNA lacking the epitope of the cytoplasmic peptide antibody. The sizes of the fragments indicated that both the intracellular and extracellular domains of t4 are proteolytically processed. To examine the processing of the/L subunit in epithelial tissues in vivo, human skin frozen sections were stained with antibodies to the ectodomain or the cytoplasmic domain of t4 The distinct staining patterns obtained with the two types of antibodies provided evidence that t4 is proteolyticaUy processed in vivo in skin. Analogous experiments performed on sections of the cornea suggested that t , is not proteolytically processed at a detectable level in this tissue. Thus, cleavage of the t4 subunit occurs in a tissue-specific fashion. These results suggest a potential mechanism of modulating the activities of the cx6fl4 integrin. T i HE Ot~4 integrin is expressed by epithelial and neuroepithelial cells (22). In epithelial tissues, this integrin is restricted to the basal portion of those cells which contact the basement membrane, suggesting a role in adhesion to basement membranes (4,12,22,24,25). Its ligand may be laminin or a laminin isoform (15, 23). The primary sequences of both the ct~ and the t, subunits have been recently determined (10, 27, 28). The t , subunit is unique among integrin fl subunits because of the large size of its cytoplasmic domain. This part of the molecule, which measures over 100 kD, contains two pairs of type III fibronectin repeats interrupted by a sequence which shows variations, probably as a result of alternative splicing of mRNA (10,27,28). Type m fibronectin repeats are commonly found in extracellular matrix molecules and in the external portion of many adhesion receptors (16). In addition to the intracytoplasmic portion of f14, two other known intracellular mole- cules with such repeats are twitchin and titin, two related proteins of the myofibrillar cytoskeleton (1)...

show abstract

Section: Discussionmentioning

confidence: 99%

Section: Immunohistochemical Detection Of B~ and Its Proteolytic Fragmentioning

confidence: 99%

See 1 more Smart Citation

Proteolytic processing of endogenous and recombinant beta 4 integrin subunit.

Giancotti

Stepp

Suzuki

et al. 1992

The Journal of Cell Biology

122

View full text Add to dashboard Cite

show abstract

“…For adhesion assays, performed as previously described, 36 96-well plates were coated with 0.1% BSA in calcium-and magnesium-free phosphate-buffered saline (PBS), 10 g/ml collagen IV (Sigma-Aldrich) in 10 mm acetic acid, 10 g/ml laminin-1 (Sigma-Aldrich) in PBS, or 10 g/ml human fibronectin (Sigma-Aldrich) overnight at 4°C. Keratinocytes were plated (30 000 cells per well) in medium in the absence of serum.…”

Section: Cell Adhesion and Migration Assaysmentioning

confidence: 99%

Corrective gene transfer of keratinocytes from patients with junctional epidermolysis bullosa restores assembly of hemidesmosomes in reconstructed epithelia

et al. 1998

Self Cite

View full text Add to dashboard Cite

Herlitz junctional epidermolysis bullosa (H-JEB) providesdeposited into the extracellular matrix. Re-expression of a promising model for somatic gene therapy of heritable laminin-5 induced cell spreading, nucleation of hemidesmechano-bullous disorders. This genodermatosis is mosomal-like structures and enhanced adhesion to the culcaused by the lack of laminin-5 that results in absence of ture substrate. Organotypic cultures performed with the hemidesmosomes (HD) and defective adhesion of squamtransduced keratinocytes, reconstituted epidermis closely ous epithelia. To establish whether re-expression of lamiadhering to the mesenchyme and presenting mature heminin-5 can restore assembly of the dermal-epidermal attachdesmosomes, bridging the cytoplasmic intermediate filament structures lacking in the H-JEB skin, we corrected the ments of the basal cells to the anchoring filaments of the genetic mutation hindering expression of the ␤3 chain of basement membrane. Our results provide the first evilaminin-5 in human H-JEB keratinocytes by transfer of a dence of phenotypic reversion of JEB keratinocytes by laminin ␤3 transgene. The transduced keratinocytes synsomatic gene therapy and demonstrate that genetic treatthesized a recombinant ␤3 polypeptide that assembled ment of the mild forms of skin blistering diseases and other with the endogenous laminin ␣3 and ␥2 chains into a bioinherited extracellular matrix pathologies is a realistic goal. logically active laminin-5 that was secreted, processed and

show abstract

“…The cytoplasmic HD plaque components, which include HD1/plectin (3) and the bullous pemphigoid antigen 1 (BP230) (4), act as linkers for elements of the cytoskeleton at the cytoplasmic surface of plasma membrane. The trans-membrane constituents of HDs, which include the ␣ 6 ␤ 4 integrin (5,6) and the bullous pemphigoid antigen 2 (BP180) (7), serve as cell receptors connecting the cell interior to extracellular matrix proteins.…”

mentioning

confidence: 99%

Gene Correction of Integrin β4-dependent Pyloric Atresia-Junctional Epidermolysis Bullosa Keratinocytes Establishes a Role for β4 Tyrosines 1422 and 1440 in Hemidesmosome Assembly

Dellambra¹,

Prislei²,

Salvati³

et al. 2001

Journal of Biological Chemistry

View full text Add to dashboard Cite

The cytoplasmic domain of ␤ 4 integrin contains two pairs of fibronectin-like repeats separated by a connecting segment. The connecting segment harbors a putative tyrosine activation motif in which tyrosines 1422 and 1440 are phosphorylated in response to ␣ 6 ␤ 4 binding to laminin-5. Primary ␤ 4 -null keratinocytes, obtained from a newborn suffering from lethal junctional epidermolysis bullosa, were stably transduced with retroviruses carrying a full-length ␤ 4 cDNA or a ␤ 4 cDNA with phenylalanine substitutions at Tyr-1422 and Tyr-1440. Hemidesmosome assembly was evaluated on organotypic skin cultures. ␤ 4 -corrected keratinocytes were indistinguishable from normal cells in terms of ␣ 6 ␤ 4 expression, the localization of hemidesmosome components, and hemidesmosome structure and density, suggesting full genetic and functional correction of ␤ 4 -null keratinocytes. In cultures generated from ␤ 4 Y1422F/Y1440Fkeratinocytes, ␤ 4 mutants as well as ␣ 6 integrin, HD1/ plectin, and BP180 were not concentrated at the dermalepidermal junction. Furthermore, the number of hemidesmosomes was strikingly reduced as compared with ␤ 4 -corrected keratinocytes. The rare hemidesmosomes detected in ␤ 4 Y1422F/Y1440F cells were devoid of subbasal dense plates and of inner cytoplasmic plaques with keratin filament insertion. Collectively, our data demonstrate that the ␤ 4 tyrosine activation motif is not required for the localization of ␣ 6 ␤ 4 at the keratinocyte plasma membrane but is essential for optimal assembly of bona fide hemidesmosomes.Human epidermis consists of a stratified squamous epithelium composed of keratinocytes organized in distinct cellular layers. Keratinocytes forming the basal layer firmly adhere to the basement membrane by means of hemidesmosomes (HDs), 1 multiprotein complexes linking the epithelial intermediate filament network to the dermal anchoring fibrils (see Refs. 1 and 2 for review). HDs are formed by the clustering of several cytoplasmic and trans-membrane proteins (2). The cytoplasmic HD plaque components, which include HD1/plectin (3) and the bullous pemphigoid antigen 1 (BP230) (4), act as linkers for elements of the cytoskeleton at the cytoplasmic surface of plasma membrane. The trans-membrane constituents of HDs, which include the ␣ 6 ␤ 4 integrin (5, 6) and the bullous pemphigoid antigen 2 (BP180) (7), serve as cell receptors connecting the cell interior to extracellular matrix proteins.In particular, the ␣ 6 ␤ 4 integrin is a receptor for the basement membrane component laminin-5, a heterotrimeric protein composed of three distinct polypeptides, ␣3, ␤3, and ␥2, which are encoded by three different genes known as LAMA3, LAMB3, and LAMC2, respectively (see Ref. 8 for review). Laminin-5 binds to the basal keratinocyte cell surface through the ␣ 6 ␤ 4 integrin and tightens the dermal-epidermal junction by binding also to the N-terminal NC-1 domain of type VII collagen (9). The crucial importance of the interaction between laminin-5 and its ␣ 6 ␤ 4 receptor in maintaining the integrity of the i...

show abstract

Polarized integrin mediates human keratinocyte adhesion to basal lamina.

Cited by 198 publications

References 35 publications

Proteolytic processing of endogenous and recombinant beta 4 integrin subunit.

Proteolytic processing of endogenous and recombinant beta 4 integrin subunit.

Corrective gene transfer of keratinocytes from patients with junctional epidermolysis bullosa restores assembly of hemidesmosomes in reconstructed epithelia

Gene Correction of Integrin β4-dependent Pyloric Atresia-Junctional Epidermolysis Bullosa Keratinocytes Establishes a Role for β4 Tyrosines 1422 and 1440 in Hemidesmosome Assembly

Contact Info

Product

Resources

About