Cellular Interaction of Integrin α3β1 with Laminin 5 Promotes Gap Junctional Communication

Lampe, Paul D.; Nguyen, Beth P.; Gil, Susana María; Usui, Marcia L.; Olerud, John E.; Takada, Yoshikazu; Carter, William G.

doi:10.1083/jcb.143.6.1735

Cited by 155 publications

(150 citation statements)

References 76 publications

(125 reference statements)

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“…In addition, both syndecan-1 protein and mRNA levels are specifically and strongly induced in wound edge keratinocytes during wound healing (48 -51). This expression resembles that of unprocessed LN-5 deposited in the provisional basement membrane by leading keratinocytes during wound healing (6,25), reinforcing our results showing its interaction with syndecan-1.…”

Section: Discussionsupporting

confidence: 74%

“…In vivo, epidermal injury activates the transcription and deposition of LN-5 into the provisional matrix by the leading keratinocytes in the process of epidermal outgrowth and migration at the wound edge (24 -26). The LN-5 precursor is found in this provisional matrix but is absent from mature basement membranes (21,25). These data indicate that the presence of the LG4/5 domain in the ␣3 chain of LN-5 may influence cell behavior.…”

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confidence: 76%

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Normal Human Keratinocytes Bind to the α3LG4/5 Domain of Unprocessed Laminin-5 through the Receptor Syndecan-1

Okamoto

Bachy

Odenthal³

et al. 2003

Journal of Biological Chemistry

101

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Basal keratinocytes of the epidermis adhere to their underlying basement membrane through a specific interaction with laminin-5, which is composed by the association of ␣3, ␤3, and ␥2 chains. Laminin-5 has the ability to induce either stable cell adhesion or migration depending on specific processing of different parts of the molecule. One event results in the cleavage of the carboxyl-terminal globular domains 4 and 5 (LG4/5) of the ␣3 chain. In this study, we recombinantly expressed the human ␣3LG4/5 fragment in mammalian cells, and we show that this fragment induces adhesion of normal human keratinocytes and fibrosarcoma-derived HT1080 cells in a heparan-and chondroitin sulfate-dependent manner. Immunoprecipitation experiments with Na 2 35 SO 4 -labeled keratinocyte and HT1080 cell lysates as well as immunoblotting experiments revealed that the major proteoglycan receptor for the ␣3LG4/5 fragment is syndecan-1. Syndecan-4 from keratinocytes also bound to ␣3LG4/5. Furthermore we could show for the first time that unprocessed laminin-5 specifically binds syndecan-1, while processed laminin-5 does not. These results demonstrate that the LG4/5 modules within unprocessed laminin-5 permit its cell binding activity through heparan and chondroitin sulfate chains of syndecan-1 and reinforce previous data suggesting specific properties for the precursor molecule.Laminins (LNs) 1 are extracellular matrix glycoproteins composed of ␣, ␤, and ␥ chains assembled into a cross-shaped heterotrimer (␣␤␥) by forming a triple-stranded coiled-coil structure through their ␣-helical domains. At present, 15 heterotrimers termed LN-1 to LN-15 have been described with different subunit composition selected from five individual ␣ chains (␣1-␣5), three ␤ chains (␤1-␤3), and three ␥ chains (␥1-␥3) (1, 2). All LN ␣ chains comprise a large globular domain in their carboxylterminal region (G domain), which consists of five homologous globular subdomains of about 200 amino acids each (LG1-LG5). LN-5, with chain composition ␣3␤3␥2, is a component of basement membranes underlying specialized epithelia with secretory or protective function (3). In skin, LN-5 is synthesized by keratinocytes initially as a high molecular mass precursor protein of 460 kDa of which the ␣3 and ␥2 chains undergo specific processing to smaller forms after being secreted and deposited into the extracellular matrix (ECM) (4, 5). Processing of the ␣3 chain consists of cleavage of the carboxyl-terminal globular domains 4 and 5 (LG4/5) (6, 7). An additional cleavage within the aminoterminal domain IIIa of the ␣3 chain occurs subsequently and may also be important for LN-5 function (8, 9). Processing of the ␥2 chain occurs at the amino terminus with loss of a 50-kDa domain (9 -11). Processed LN-5 is the major component of anchoring filaments in skin (12) where it mediates cell adhesion via interaction of the ␣3 carboxyl-terminal LG1-3 triplet domain with both ␣ 3 ␤ 1 and ␣ 6 ␤ 4 integrins (13-16). Several studies have suggested that processed LN-5 functions both in the nuclea...

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Section: Discussionsupporting

confidence: 74%

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confidence: 76%

Normal Human Keratinocytes Bind to the α3LG4/5 Domain of Unprocessed Laminin-5 through the Receptor Syndecan-1

Okamoto

Bachy

Odenthal³

et al. 2003

Journal of Biological Chemistry

101

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“…By contrast, several of the candidate proteins identified in the screen are more likely to regulate connexins through changes in cell signaling or differentiated state. For instance, although decorin B has not been shown to regulate intercellular communication, cross-talk between integrins, other extracellular matrix proteins, and connexins has previously been demonstrated (Isakson et al 2001;Kalra et al 2006;Lampe et al 1998). A similar case could be made for FRG1, which is associated with an autosomal dominant disease, facioscapulohumeral muscular dystrophy (Gabellini et al 2006).…”

Section: Discussionmentioning

confidence: 88%

Identification of rab20 as a Potential Regulator of Connexin43 Trafficking

Sarma

Kaplan

Willemsen

et al. 2008

Cell Communication & Adhesion

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Connexin oligomerization and trafficking are regulated processes. To identify proteins that control connexin43 (Cx43), a screen was designed using HeLa cells expressing a Cx43 construct with di-lysine endoplasmic reticulum (ER)-retention/retrieval motif, Cx43-HKKSL. At moderate levels of expression, Cx43-HKKSL is retained in the ER as monomers; however, Cx43-HKKSL stably overexpressed by HeLa cells localizes to the perinuclear region and oligomerizes. HeLa/Cx43-HKKSL overexpressors were transiently transfected with pooled clones from a human kidney cDNA library and used immunofluorescence microscopy to identify cDNAs that enabled overexpressed Cx43-HKKSL to convert from a perinuclear to ER localization pattern. Using this approach, a small molecular weight GTPase, rab20, was identified as a candidate protein with the ability to regulate Cx43 trafficking. Enhanced green florescent protein (EGFP)-tagged rab20 showed a predominantly perinuclear and ER localization pattern and caused wild-type Cx43 to be retained inside the cell. By contrast, mutant EGFP-rab20T19N, which lacks the ability to bind GTP, had no effect on Cx43. These results suggest Cx43 is transported through an intracellular compartment regulated by rab20 along the secretory pathway.

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“…Quiescent epidermis adheres to laminin 5 in the BM via integrin ␣ 6 ␤ 4 in hemidesmosome (HD) cell junctions. Wounding epidermis generates leading and following subpopulations of keratinocytes at the wound margin (5). Leading keratinocytes migrate over exposed dermal collagen via integrin ␣ 2 ␤ 1 and fibronectin via integrin ␣ 5 ␤ 1 .…”

mentioning

confidence: 99%

Adhesion or Plasmin Regulates Tyrosine Phosphorylation of a Novel Membrane Glycoprotein p80/gp140/CUB Domain-containing Protein 1 in Epithelia

Brown

Yang

Zaitsevskaia

et al. 2004

Journal of Biological Chemistry

Self Cite

138

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Suspension of cultured human foreskin keratinocytes (HKs) with trypsin phosphorylates tyrosine residues on an 80-kDa membrane glycoprotein, p80 (Xia, Y., Gil, S. G., and Carter, W. G. (1996) J. Cell Biol. 132, 727-740). Readhesion dephosphorylates p80. Sequencing of a p80 cDNA established identity to CUB domain-containing protein 1 (CDCP1), a gene elevated in carcinomas. CDCP1/p80 cDNA encodes three extracellular CUB domains, a transmembrane domain, and two putative cytoplasmic Tyr phosphorylation sites. Treatment of adherent HKs with suramin, a heparin analogue, or inhibitors of phosphotyrosine phosphatases (PTPs; vanadate or calpeptin) increases phosphorylation of p80 and a novel 140-kDa membrane glycoprotein, gp140. Phosphorylated gp140 was identified as a trypsin-sensitive precursor to p80. Identity was confirmed by digestion and phosphorylation studies with recombinant gp140-GFP. Plasmin, a serum protease, also converts gp140 to p80, providing biological significance to the cleavage in wounds. Phosphorylation of gp140 and p80 are mediated by Src family kinases at multiple Tyr residues including Tyr 734 . Dephosphorylation is mediated by PTP(s). Conversion of gp140 to p80 prolongs phosphorylation of p80 in response to suramin and changes in adhesion. This distinguishes gp140 and p80 and explains the relative abundance of phosphorylated p80 in trypsinized HKs. We conclude that phosphorylation of gp140 is dynamic and balanced by Src family kinase and PTPs yielding low equilibrium phosphorylation. We suggest that the balance is altered by conversion of gp140 to p80 and by adhesion, providing a novel transmembrane phosphorylation signal in epithelial wounds.Wounding of quiescent epidermis activates changes in adhesion and cell signaling resulting in keratinocyte migration, basement membrane (BM) 1 repair, and wound closure (1-3).Based on an in vitro model for wound activation, we reported that trypsin detachment of cultured human foreskin keratinocytes (HKs) promotes phosphorylation of tyrosine residues on an 80-kDa membrane glycoprotein (p80) (4). Phosphorylated p80 (P-p80) in suspended HKs is dephosphorylated upon readhesion to laminin 5 via integrins ␣ 6 ␤ 4 and ␣ 3 ␤ 1 . In work here, we purified and characterized p80. We wished to understand whether phosphorylated p80 identified in an in vitro deadhesion/readhesion screen (4) might be involved in physiologically significant wound activation. Quiescent epidermis adheres to laminin 5 in the BM via integrin ␣ 6 ␤ 4 in hemidesmosome (HD) cell junctions. Wounding epidermis generates leading and following subpopulations of keratinocytes at the wound margin (5). Leading keratinocytes migrate over exposed dermal collagen via integrin ␣ 2 ␤ 1 and fibronectin via integrin ␣ 5 ␤ 1 . However, leading cells also deposit laminin 5 as a provisional BM and interact with these deposits via integrin ␣ 3 ␤ 1 (5-7). The interaction of leading cells with deposited laminin 5 generates distinct transmembrane signals when compared with interaction with dermal ligands. For example,...

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Cellular Interaction of Integrin α3β1 with Laminin 5 Promotes Gap Junctional Communication

Cited by 155 publications

References 76 publications

Normal Human Keratinocytes Bind to the α3LG4/5 Domain of Unprocessed Laminin-5 through the Receptor Syndecan-1

Normal Human Keratinocytes Bind to the α3LG4/5 Domain of Unprocessed Laminin-5 through the Receptor Syndecan-1

Identification of rab20 as a Potential Regulator of Connexin43 Trafficking

Adhesion or Plasmin Regulates Tyrosine Phosphorylation of a Novel Membrane Glycoprotein p80/gp140/CUB Domain-containing Protein 1 in Epithelia

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