scholarly journals Induction of tyrosine phosphorylation during ICAM-3 and LFA-1-mediated intercellular adhesion, and its regulation by the CD45 tyrosine phosphatase.

1994 ◽  
Vol 126 (5) ◽  
pp. 1277-1286 ◽  
Author(s):  
A G Arroyo ◽  
M R Campanero ◽  
P Sánchez-Mateos ◽  
J M Zapata ◽  
M A Ursa ◽  
...  
Keyword(s):  
Cell Adhesion ◽  
Cell Line ◽  
Tyrosine Phosphorylation ◽  
Molecular Mechanisms ◽  
Tyrosine Phosphatase ◽  
Cell Aggregation ◽  
Intercellular Adhesion ◽  
Lymphocyte Function ◽  
Late Antigen ◽  
Cell Cell

Intercellular adhesion molecule (ICAM)-3, a recently described counter-receptor for the lymphocyte function-associated antigen (LFA)-1 integrin, appears to play an important role in the initial phase of immune response. We have previously described the involvement of ICAM-3 in the regulation of LFA-1/ICAM-1-dependent cell-cell interaction of T lymphoblasts. In this study, we further investigated the functional role of ICAM-3 in other leukocyte cell-cell interactions as well as the molecular mechanisms regulating these processes. We have found that ICAM-3 is also able to mediate LFA-1/ICAM-1-independent cell aggregation of the leukemic JM T cell line and the LFA-1/CD18-deficient HAFSA B cell line. The ICAM-3-induced cell aggregation of JM and HAFSA cells was not affected by the addition of blocking mAb specific for a number of cell adhesion molecules such as CD1 1a/CD18, ICAM-1 (CD54), CD2, LFA-3 (CD58), very late antigen alpha 4 (CD49d), and very late antigen beta 1 (CD29). Interestingly, some mAb against the leukocyte tyrosine phosphatase CD45 were able to inhibit this interaction. Moreover, they also prevented the aggregation induced on JM T cells by the proaggregatory anti-LFA-1 alpha NKI-L16 mAb. In addition, inhibitors of tyrosine kinase activity also abolished ICAM-3 and LFA-1-mediated cell aggregation. The induction of tyrosine phosphorylation through ICAM-3 and LFA-1 antigens was studied by immunofluorescence, and it was found that tyrosine-phosphorylated proteins were preferentially located at intercellular boundaries upon the induction of cell aggregation by either anti-ICAM-3 or anti-LFA-1 alpha mAb. Western blot analysis revealed that the engagement of ICAM-3 or LFA-1 with activating mAb enhanced tyrosine phosphorylation of polypeptides of 125, 70, and 38 kD on JM cells. This phenomenon was inhibited by preincubation of JM cells with those anti-CD45 mAb that prevented cell aggregation. Altogether these results indicate that CD45 tyrosine phosphatase plays a relevant role in the regulation of both intracellular signaling and cell adhesion induced through ICAM-3 and beta 2 integrins.

scholarly journals ICAM-3 interacts with LFA-1 and regulates the LFA-1/ICAM-1 cell adhesion pathway.

1993 ◽  
Vol 123 (4) ◽  
pp. 1007-1016 ◽  
Author(s):  
M R Campanero ◽  
M A del Pozo ◽  
A G Arroyo ◽  
P Sánchez-Mateos ◽  
T Hernández-Caselles ◽  
...  
Keyword(s):  
Cell Adhesion ◽  
T Cell ◽  
T Cell Activation ◽  
Cell Aggregation ◽  
Intercellular Adhesion ◽  
Intercellular Adhesion Molecule ◽  
Lymphocyte Function ◽  
T Lymphocyte Proliferation ◽  
Cell Cell ◽  
T Cell Adhesion

The interaction of lymphocyte function-associated antigen-1 (LFA-1) with its ligands mediates multiple cell adhesion processes of capital importance during immune responses. We have obtained three anti-ICAM-3 mAbs which recognize two different epitopes (A and B) on the intercellular adhesion molecule-3 (ICAM-3) as demonstrated by sequential immunoprecipitation and cross-competitive mAb-binding experiments. Immunoaffinity purified ICAM-3-coated surfaces were able to support T lymphoblast attachment upon cell stimulation with both phorbol esters and cross-linked CD3, as well as by mAb engagement of the LFA-1 molecule with the activating anti-LFA-1 NKI-L16 mAb. T cell adhesion to purified ICAM-3 was completely inhibited by cell pretreatment with mAbs to the LFA-1 alpha (CD11a) or the LFA-beta (CD18) integrin chains. Anti-ICAM-3 mAbs specific for epitope A, but not those specific for epitope B, were able to trigger T lymphoblast homotypic aggregation. ICAM-3-mediated cell aggregation was dependent on the LFA-1/ICAM-1 pathway as demonstrated by blocking experiments with mAbs specific for the LFA-1 and ICAM-1 molecules. Furthermore, immunofluorescence studies on ICAM-3-induced cell aggregates revealed that both LFA-1 and ICAM-1 were mainly located at intercellular boundaries. ICAM-3 was located at cellular uropods, which in small aggregates appeared to be implicated in cell-cell contacts, whereas in large aggregates it appeared to be excluded from cell-cell contact areas. Experiments of T cell adhesion to a chimeric ICAM-1-Fc molecule revealed that the proaggregatory anti-ICAM-3 HP2/19 mAb was able to increase T lymphoblast attachment to ICAM-1, suggesting that T cell aggregation induced by this mAb could be mediated by increasing the avidity of LFA-1 for ICAM-1. Moreover, the HP2/19 mAb was costimulatory with anti-CD3 mAb for T lymphocyte proliferation, indicating that enhancement of T cell activation could be involved in ICAM-3-mediated adhesive phenomena. Altogether, our results indicate that ICAM-3 has a regulatory role on the LFA-1/ICAM-1 pathway of intercellular adhesion.

scholarly journals Cadherin-mediated cell-cell adhesion is perturbed by v-src tyrosine phosphorylation in metastatic fibroblasts.

1992 ◽  
Vol 118 (3) ◽  
pp. 703-714 ◽  
Author(s):  
N Matsuyoshi ◽  
M Hamaguchi ◽  
S Taniguchi ◽  
A Nagafuchi ◽  
S Tsukita ◽  
...  
Keyword(s):  
Cell Adhesion ◽  
Tyrosine Phosphorylation ◽  
Kinase Inhibitor ◽  
Tyrosine Phosphatase ◽  
Metastatic Potential ◽  
Transformed Cells ◽  
Adhesive Properties ◽  
Herbimycin A ◽  
Associated Proteins ◽  
Cell Cell

Rat 3Y1 cells acquire metastatic potential when transformed with v-src, and this potential is enhanced by double transformation with v-src and v-fos (Taniguchi, S., T. Kawano, T. Mitsudomi, G. Kimura, and T. Baba. 1986. Jpn. J. Cancer Res. 77:1193-1197). We compared the activity of cadherin cell adhesion molecules of normal 3Y1 cells with that of v-src transformed (SR3Y1) and v-src and v-fos double transformed (fosSR3Y1) 3Y1 cells. These cells expressed similar amounts of P-cadherin, and showed similar rates of cadherin-mediated aggregation under suspended conditions. However, the aggregates or colonies of these cells were morphologically distinct. Normal 3Y1 cells formed compacted aggregates in which cells are firmly connected with each other, whereas the transformed cells were more loosely associated, and could freely migrate out of the colonies. Overexpression of exogenous E-cadherin in these transformed cells had no significant effect on their adhesive properties. We then found that herbimycin A, a tyrosine kinase inhibitor, induced tighter cell-cell associations in the aggregates of the transformed cells. In contrast, vanadate, a tyrosine phosphatase inhibitor, inhibited the cadherin-mediated aggregation of SR3Y1 and fosSR3Y1 cells but had little effect on that of normal 3Y1 cells. These results suggest that v-src-mediated tyrosine phosphorylation perturbs cadherin function directly or indirectly, and the inhibition of tyrosine phosphorylation restores cadherin action to the normal state. We next studied tyrosine phosphorylation on cadherins and the cadherin-associated proteins, catenins. While similar amounts of catenins were expressed in all of these cells, the 98-kD catenin was strongly tyrosine phosphorylated only in SR3Y1 and fosSR3Y1 cells. Cadherins were also weakly tyrosine phosphorylated only in the transformed cells. The tyrosine phosphorylation of these proteins was enhanced by vanadate, and inhibited by herbimycin A. Thus, the tyrosine phosphorylation of the cadherin-catenin system itself might affect its function, causing instable cell-cell adhesion.

scholarly journals The Protein Tyrosine Phosphatase Pez Is a Major Phosphatase of Adherens Junctions and Dephosphorylates β-Catenin

2003 ◽  
Vol 14 (6) ◽  
pp. 2520-2529 ◽  
Author(s):  
Carol Wadham ◽  
Jennifer R Gamble ◽  
Mathew A Vadas ◽  
Yeesim Khew-Goodall
Keyword(s):  
Cell Adhesion ◽  
Tyrosine Phosphorylation ◽  
Adherens Junctions ◽  
Tyrosine Phosphatase ◽  
Adherens Junction ◽  
Dominant Negative ◽  
Dominant Negative Mutant ◽  
Adherens Junction Proteins ◽  
Cell Cell

Cell-cell adhesion regulates processes important in embryonal development, normal physiology, and cancer progression. It is regulated by various mechanisms including tyrosine phosphorylation. We have previously shown that the protein tyrosine phosphatase Pez is concentrated at intercellular junctions in confluent, quiescent monolayers but is nuclear in cells lacking cell-cell contacts. We show here with an epithelial cell model that Pez localizes to the adherens junctions in confluent monolayers. A truncation mutant lacking the catalytic domain acts as a dominant negative mutant to upregulate tyrosine phosphorylation at adherens junctions. We identified β-catenin, a component of adherens junctions, as a substrate of Pez by a “substrate trapping” approach and by in vitro dephosphorylation with recombinant Pez. Consistent with this, ectopic expression of the dominant negative mutant caused an increase in tyrosine phosphorylation of β-catenin, demonstrating that Pez regulates the level of tyrosine phosphorylation of adherens junction proteins, including β-catenin. Increased tyrosine phosphorylation of adherens junction proteins has been shown to decrease cell-cell adhesion, promoting cell migration as a result. Accordingly, the dominant negative Pez mutant enhanced cell motility in an in vitro “wound” assay. This suggests that Pez is also a regulator of cell motility, most likely through its action on cell-cell adhesion.

B-cell homotypic adhesion through exon-A restricted epitopes of CD45 involves LFA-1/ICAM-1, ICAM-3 interactions, and induces coclustering of CD45 and LFA-1

Blood ◽  
1995 ◽  
Vol 86 (5) ◽  
pp. 1861-1872 ◽  
Author(s):  
JM Zapata ◽  
MR Campanero ◽  
M Marazuela ◽  
F Sanchez-Madrid ◽  
MO de Landazuri
Keyword(s):  
Cell Adhesion ◽  
B Cell ◽  
Phosphatase Activity ◽  
Peripheral Blood ◽  
Membrane Surface ◽  
Cell Aggregation ◽  
Intercellular Adhesion ◽  
Cell Contact ◽  
Phosphotyrosine Phosphatase ◽  
Cell Cell

Lymphocyte interactions with other leukocytes and other cell types, as well as with components of the extracellular matrix, are one of the key steps in the immune response. Three novel monoclonal antibodies (MoAbs) have been produced and selected for their ability to induce intercellular adhesion in B cells. These three MoAbs immunoprecipitated a polypeptide of 220 kD, displaying specific phosphotyrosine phosphatase activity that has been identified as CD45. These MoAbs recognize epitopes located on the alternative spliced exon-A-encoded region of CD45. These epitopes are of polypeptidic nature, but they can be masked by addition of carbohydrate during CD45 biosynthesis. Interestingly enough, CD45 epitopes recognized by these MoAbs appeared to be selectively expressed on both peripheral blood and tonsillar B lymphocytes as well as on peripheral blood natural killer (NK) cells. CD45-mediated intercellular adhesion was abrogated upon incubation with anti-leukocyte function-associated antigen 1 (anti-LFA-1), intercellular cell adhesion molecule 1 (ICAM-1), and ICAM-3 MoAbs, thus indicating that this phenomenon involved both LFA-1/ICAM-1 and LFA- 1/ICAM-3 cell adhesion pathways. Moreover, CD45-mediated cell aggregation was also inhibited by preincubation with some conventional anti-CD45 MoAbs. Interestingly, the triggering of cell aggregation through CD45 induced membrane surface relocation of CD45 and LFA-1 molecules, with both of them colocalizing at cell-cell contact areas of B-cell aggregates. Studies with inhibitors of both phosphotyrosine phosphatase and tyrosine kinase activities suggest that CD45 phosphotyrosine phosphatase activity could be involved in CD45-mediated cell aggregation. Taken together, these results support the notion that CD45 is a key molecule in the regulation of LFA-1-mediated cell-cell interactions.

scholarly journals Loss of E-cadherin-dependent cell-cell adhesion due to mutation of the beta-catenin gene in a human cancer cell line, HSC-39.

1995 ◽  
Vol 15 (3) ◽  
pp. 1175-1181 ◽  
Author(s):  
J Kawanishi ◽  
J Kato ◽  
K Sasaki ◽  
S Fujii ◽  
N Watanabe ◽  
...  
Keyword(s):  
Cell Adhesion ◽  
Cell Line ◽  
Cancer Cell ◽  
Cancer Cell Line ◽  
Cell Aggregation ◽  
Expression Vectors ◽  
Beta Catenin ◽  
Dependent Cell ◽  
E Cadherin ◽  
Cell Cell

Detachment of cell-cell adhesion is indispensable for the first step of invasion and metastasis of cancer. This mechanism is frequently associated with the impairment of either E-cadherin expression or function. However, mechanisms of such abnormalities have not been fully elucidated. In this study, we demonstrated that the function of E-cadherin was completely abolished in the human gastric cancer cell line HSC-39, despite the high expression of E-cadherin, because of mutations in one of the E-cadherin-associated cytoplasmic proteins, beta-catenin. Although immunofluorescence staining of HSC-39 cells by using an anti-E-cadherin antibody (HECD-1) revealed the strong and uniform expression of E-cadherin on the cell surface, cell compaction and cell aggregation were not observed in this cell. Western blotting (immunoblotting) using HECD-1 exhibited a 120-kDa band which is equivalent to normal E-cadherin. Northern (RNA) blotting demonstrated a 4.7-kb band, the same as mature E-cadherin mRNA. Immunoprecipitation of metabolically labeled proteins with HECD-1 revealed three bands corresponding to E-cadherin, alpha-catenin, and gamma-catenin and a 79-kDa band which was apparently smaller than that of normal beta-catenin, indicating truncated beta-catenin. The 79-kDa band was immunologically identified as beta-catenin by using immunoblotting with anti-beta-catenin antibodies. Examination of beta-catenin mRNA by the reverse transcriptase-PCR method revealed a transcript which was shorter than that of normal beta-catenin. The sequencing of PCR product for beta-catenin confirmed deletion in 321 bases from nucleotides +82 to +402. Southern blotting of beta-catenin DNA disclosed mutation at the genomic level. Expression vectors of Beta-catenin were introduced into HSC-39 cells by transfection. In the obtained transfectants, E-cadherin-dependent cell-cell adhesiveness was recovered, as revealed by cell compaction, cell aggregation, and immunoflourescence staining. From these results, it was concluded that in HSC-39 cells, impaired cell-cell adhesion is due to mutations in beta-catenin which results in the dysfunction of E-cadherin.

Cell Aggregation and Tissue Equivalents

2004 ◽  
Author(s):  
W. Mark Saltzman
Keyword(s):  
Cell Adhesion ◽  
Information Exchange ◽  
Molecular Mechanisms ◽  
Biological Systems ◽  
Cell Aggregation ◽  
Dynamic Interactions ◽  
Cell Shape Change ◽  
Aggregation Pattern ◽  
Other Information ◽  
Cell Cell

Development of an individual from a fertilized egg involves a rich choreography of cell division, cell movement, and cell shape change. Cells replicate, interact with their surroundings, and move individually and in coordination. Development produces a multicellular organism in which cells are organized into cooperative communities. Adult tissues and organs are orderly ensembles of cells—usually the ensemble contains more than one type of cell—in which both ensemble architecture and dynamic interactions between individual cells are critical for proper function. Tissues are collections of cells, united for a common function. Tissues that differ in function also differ in architecture, that is, in the shape and arrangement of cells. Even in the absence of other information, this observation could lead one to speculate on the power of the influence of cell–cell interactions on tissue development and function. It is well known that selective cell adhesion is a fundamental mechanism underlying mammalian development (see Chapter 3). But cell adhesion and cell–cell communication are essential throughout the lifetime of an organism. The previous three chapters described the biophysical processes of division, adhesion, and motility and introduced some of the molecular mechanisms that cells use to interact with, and learn about, their environment. This chapter considers the relationship of cell adhesion to cell aggregation, or the formation and maintenance of cell communities. The role of cell adhesion in circulation and trafficking of cells throughout the body is discussed in Chapter 10. A community is often more effective than the sum of its members. This is abundantly evident in biological systems; swarms of bees and colonies of ants are more effective than their numbers suggest because bees and ants can act in coordination. Synergistic activity in multicomponent biological systems requires information exchange between units and control mechanisms to guide a unit response. Cell aggregates are communities of cells that are organized in a specific three-dimensional architecture. The preferred aggregation pattern within a particular tissue or organ has important consequences with regard to tissue function.

scholarly journals Candida albicans Expresses a Focal Adhesion Kinase-Like Protein That Undergoes Increased Tyrosine Phosphorylation upon Yeast Cell Adhesion to Vitronectin and the EA.hy 926 Human Endothelial Cell Line

2002 ◽  
Vol 70 (7) ◽  
pp. 3804-3815 ◽  
Author(s):  
Giorgio Santoni ◽  
Roberta Lucciarini ◽  
Consuelo Amantini ◽  
Jordan Jacobelli ◽  
Elisabetta Spreghini ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Candida Albicans ◽  
Cell Adhesion ◽  
Endothelial Cell ◽  
Yeast Cell ◽  
Focal Adhesion Kinase ◽  
Cell Line ◽  
Tyrosine Phosphorylation ◽  
Focal Adhesion ◽  
Human Endothelial Cell Line

ABSTRACT The signaling pathways triggered by adherence of Candida albicans to the host cells or extracellular matrix are poorly understood. We provide here evidence in C. albicans yeasts of a p105 focal adhesion kinase (Fak)-like protein (that we termed CaFak), antigenically related to the vertebrate p125Fak, and its involvement in integrin-like-mediated fungus adhesion to vitronectin (VN) and EA.hy 926 human endothelial cell line. Biochemical analysis with different anti-chicken Fak antibodies identified CaFak as a 105-kDa protein and immunofluorescence and cytofluorimetric analysis on permeabilized cells specifically stain C. albicans yeasts; moreover, confocal microscopy evidences CaFak as a cytosolic protein that colocalizes on the membrane with the integrin-like VN receptors upon yeast adhesion to VN. The protein tyrosine kinase (PTK) inhibitors genistein and herbimycin A strongly inhibited C. albicans yeast adhesion to VN and EA.hy 926 endothelial cells. Moreover, engagement of αvβ3 and αvβ5 integrin-like on C. albicans either by specific monoclonal antibodies or upon adhesion to VN or EA.hy 926 endothelial cells stimulates CaFak tyrosine phosphorylation that is blocked by PTK inhibitor. A role for CaFak in C. albicans yeast adhesion was also supported by the failure of VN to stimulate its tyrosine phosphorylation in a C. albicans mutant showing normal levels of CaFak and VNR-like integrins but displaying reduced adhesiveness to VN and EA.hy 926 endothelial cells. Our results suggest that C. albicans Fak-like protein is involved in the control of yeast cell adhesion to VN and endothelial cells.

CELL BEHAVIOUR AND MOLECULAR MECHANISMS OF CELL-CELL ADHESION

1981 ◽  
Vol 56 (2) ◽  
pp. 199-240 ◽  
Author(s):  
D. R. GARROD ◽  
A. NICOL
Keyword(s):  
Cell Adhesion ◽  
Molecular Mechanisms ◽  
Cell Behaviour ◽  
Cell Cell

scholarly journals Reevaluating αE-catenin monomer and homodimer functions by characterizing E-cadherin/αE-catenin chimeras

2015 ◽  
Vol 210 (7) ◽  
pp. 1065-1074 ◽  
Author(s):  
Julie M. Bianchini ◽  
Khameeka N. Kitt ◽  
Martijn Gloerich ◽  
Sabine Pokutta ◽  
William I. Weis ◽  
...  
Keyword(s):  
Cell Adhesion ◽  
Intercellular Adhesion ◽  
Dependent Cell ◽  
In Cells ◽  
E Cadherin ◽  
Cell Cell

As part of the E-cadherin–β-catenin–αE-catenin complex (CCC), mammalian αE-catenin binds F-actin weakly in the absence of force, whereas cytosolic αE-catenin forms a homodimer that interacts more strongly with F-actin. It has been concluded that cytosolic αE-catenin homodimer is not important for intercellular adhesion because E-cadherin/αE-catenin chimeras thought to mimic the CCC are sufficient to induce cell–cell adhesion. We show that, unlike αE-catenin in the CCC, these chimeras homodimerize, bind F-actin strongly, and inhibit the Arp2/3 complex, all of which are properties of the αE-catenin homodimer. To more accurately mimic the junctional CCC, we designed a constitutively monomeric chimera, and show that E-cadherin–dependent cell adhesion is weaker in cells expressing this chimera compared with cells in which αE-catenin homodimers are present. Our results demonstrate that E-cadherin/αE-catenin chimeras used previously do not mimic αE-catenin in the native CCC, and imply that both CCC-bound monomer and cytosolic homodimer αE-catenin are required for strong cell–cell adhesion.

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