Identification of the Integrin VLA-2 as a Receptor for Echovirus 1

Bergelson, Jeffrey M.; Shepley, Michael P.; Chan, Bosco M.C.; Hemler, Martin E.; Finberg, Robert W.

doi:10.1126/science.1553561

Cited by 288 publications

(178 citation statements)

References 31 publications

Supporting

Mentioning

169

Contrasting

Unclassified

Order By: Relevance

“…When grown to A 595 ϭ 0.8, the cells (800 ml) were pelleted by centrifugation, washed with M9 minimal medium (12.8 g of Na 2 HPO 4 /liter, 3 g of KH 2 PO 4 /liter, 0.5 g of NaCl/ liter, 1 g of NH 4 Cl/liter, 1 mM MgCl 2 , 0.1 mM CaCl 2 , 0.5% glucose, 2 mg of thiamine/liter), and pelleted again. The pellet was resuspended in 500 ml of M9 minimal medium supplemented with 150 mg/liter of each of the following amino acids: threonine, phenylalanine, leucine, isoleucine, lysine, valine, and proline.…”

Section: Methodsmentioning

confidence: 99%

Mapping the Collagen-binding Site in the I Domain of the Glycoprotein Ia/IIa (Integrin α2β1)

Smith¹,

Estavillo²,

Emsley³

et al. 2000

Journal of Biological Chemistry

View full text Add to dashboard Cite

The I domain present within the ␣2 chain of the integrin ␣ 2 ␤ 1 (GPIa/IIa) contains the principal collagenbinding site. Based on the crystal structure of the ␣2-I domain, a hypothetical model was proposed in which collagen binds to a groove on the upper surface of the I domain (Emsley, J., King, S. L., Bergelson, J. M., and Liddington, R. C. (1997) J. Biol. Chem. 272, 28512-28517). We have introduced point mutations into 13 residues on the upper surface of the domain. Recombinant mutant proteins were assayed for binding to monoclonal antibodies 6F1 and 12F1, to collagen under static conditions, and for the ability to retain adhesive activity under flow conditions. The mutations to residues surrounding the metal ion-dependent adhesion site that caused the greatest loss of collagen binding under both static and flow conditions are N154S in the ␤A-␣1 turn, N190D in the ␤B-␤C turn, D219R in the ␣3-␣4 turn, and E256V and H258V in the ␤D-␣5 turn. Mutation in one of the residues that coordinate the metal binding, S155A, completely lost the adhesive activity under flow but bound normally under static conditions, whereas the mutation Y285F had the converse effect. We conclude that the upper surface of the domain, including the metal ion-dependent adhesion site motif, defines the collagen recognition site. The glycoprotein (GP)1 Ia/IIa (integrin ␣ 2 ␤ 1 ) is a major platelet adhesion receptor for collagen. Integrins are a family of heterodimeric cell adhesion molecules that mediate cell-cell and cell-matrix adhesion (1). The integrin ␣ 2 ␤ 1 is expressed on several different cell types. Although it is a collagen receptor in platelet and fibroblastic cells, it functions both as a collagen and laminin receptor on endothelial and epithelial cells (2, 3). It also acts as the receptor for the human pathogen echovirus-1 (4).␣ 2 ␤ 1 is composed of a 150-kDa ␣2 and a 130-kDa ␤1 subunit (5). Within the ␣2 subunit, the 200-amino acid I domain shares homology with the A domains of von Willebrand factor, complement proteins, cartilage matrix protein, and certain other integrins. There is increasing evidence that I (A) domains play an important role in cell-adhesion protein and cell-matrix interaction.Within the ␣ 2 ␤ 1 integrin, the I domain (amino acids 140 -349) provides the principal binding site for collagen. Thus, antibodies that block ␣ 2 ␤ 1 interaction with collagen recognize epitopes within the I domain (6 -8). Three groups have shown that recombinant ␣2-I domain fusion proteins bind specifically to collagen in a divalent cation-dependent manner (9 -11), and a fourth group has presented conflicting data suggesting that collagen binding is independent of divalent cation (12).Like other I domains, the ␣2-I domain contains a cationbinding site called the metal ion-dependent adhesion site (MI-DAS) motif (13). In the crystal structure (14), the side chains of residues Ser-153, Ser-155, and Asp-254 directly coordinate a Mg 2ϩ ion, and Asp-151 and Thr-221 provide water-mediated bonds. One study reported that mutation of residues A...

show abstract

Section: Methodsmentioning

confidence: 99%

Mapping the Collagen-binding Site in the I Domain of the Glycoprotein Ia/IIa (Integrin α2β1)

Smith¹,

Estavillo²,

Emsley³

et al. 2000

Journal of Biological Chemistry

View full text Add to dashboard Cite

show abstract

“…However, the entry routes of SV40 and EV1 seem not to be identical (Marjomäki et al, 2002) and EV1 was found to be a unique tool to study the cell biology of caveolae. Importantly, EV1 uses ␣2␤1 integrin, a collagen receptor, to bind to cell surface (Bergelson et al, 1992) and therefore, the entry of EV1 can be used to study the role of integrins in caveolae function. Previous studies have shown that integrins can be coprecipitated with caveolin-1 (Wary et al, 1996(Wary et al, , 1998, suggesting that caveolae form a platform for integrin-mediated signaling.…”

Section: Introductionmentioning

confidence: 99%

Clustering Induces a Lateral Redistribution of α2β1 Integrin from Membrane Rafts to Caveolae and Subsequent Protein Kinase C-dependent Internalization

Upla

Marjomäki

Kankaanpää

et al. 2004

MBoC

151

186

View full text Add to dashboard Cite

Integrin ␣2␤1 mediates the binding of several epithelial and mesenchymal cell types to collagen. The composition of the surrounding plasma membrane, especially caveolin-1-and cholesterol-containing membrane structures called caveolae, may be important to integrin signaling. On cell surface ␣2␤1 integrin was located in the raft like membrane domain, rich in GPI-anchored proteins, rather than in caveolae. However, when antibodies were used to generate clusters of ␣2␤1 integrin, they started to move laterally on cell surface along actin filaments. During the lateral movement small clusters fused together. Finally ␣2␤1 integrin was found inside caveolae and subsequently internalized into caveosome-like perinuclear structures. The internalization process, unlike cluster formation or lateral redistribution, was dependent on protein kinase C␣ activity. Caveolae are known to be highly immobile structures and ␣2␤1 integrin clusters represent a previously unknown mechanism to activate endocytic trafficking via caveolae. The process was specific to ␣2␤1 integrin, because the antibody-mediated formation of ␣V integrin clusters activated their internalization in coated vesicles and early endosomes. In addition to natural ligands human echovirus-1 (EV1) gains entry into the cell by binding to ␣2␤1 and taking advantage of ␣2␤1 internalization via caveolae. INTRODUCTIONCaveolae are cave-like invaginations of the cell surface (for reviews see Kurzchalia and Parton, 1999;Parton 2003). They have been described as highly immobile and not involved in constitutive endocytic trafficking (Thomsen et al., 2002). Their internalization can be activated, for example, by the phosphatase inhibitor okadaic acid (Parton et al., 1994). In endothelial cells the interaction of albumin docking protein gp60 and caveolin-1 initiates the endocytosis of caveolae (Minshall et al., 2000). Caveolins are membrane proteins that are molecular markers of caveolae and shown to be crucial for the formation of these structures (Fra et al., 1995). Caveolins can be associated with cell surface growth factor receptors (Couet et al., 1997) and cell adhesion receptors (Wary et al., 1996(Wary et al., , 1998Wei et al., 1999) and caveolae might therefore play a role in cellular signaling. In addition, caveolae have been suggested to participate in the uptake of folate by pinocytosis, but this is still under discussion (Parton, 2003). Caveolin-deficient mice have given novel insight into the function of caveolae. Caveolin-1 gene knockout leads to pulmonary defects, vasoconstriction, or dilatation abnormalities and resistance to diet-induced obesity (Drab et al., 2001;Razani et al., 2001;Schubert et al., 2001;Sotgia et al., 2002;Razani et al., 2002). Simian virus 40 (SV40) has been shown to be internalized through caveolae and detailed studies focused on the virus entry mechanism have produced a lot of new information about the biology of caveolae (Pelkmans et al., 2001. We have recently shown that human echovirus 1 (EV1) is another virus using caveolae in its entry (Mar...

show abstract

“…contain the conserved arginine-glycine-aspartic acid Integrin-mediated cell entry is exploited by a number of (RGD) motif 9 and small peptides containing an RGD intracellular pathogens including adenovirus, 1,2 echodomain, particularly cyclic peptides, have high affinities virus, 3 foot-and-mouth disease virus 4 and the enterobacfor integrins. [10][11][12][13] Such peptides are easy to synthesise in terium Yersinia pseudotuberculosis.…”

Section: Introductionmentioning

confidence: 99%

Integrin-mediated transfection with peptides containing arginine-glycine-aspartic acid domains

et al. 1997

View full text Add to dashboard Cite

Two synthetic peptides comprising an RGD moiety for intecase of the COS-7 and A375M cell lines. With the ECV304 grin binding and a polylysine moiety for DNA binding were cell line, optimum transfection occurred in the absence of tested for transfection efficiency under a variety of different chloroquine. Transfection efficiency of the peptides was conditions. Binding of target cells to the peptide was shown greatest at peptide:DNA ratios of 4:1 (w/w), which were to be strongly dependent on cyclisation of the peptides via calculated to generate complexes containing approxicysteine residues. Low (10 M) concentrations of chloromately 5000 peptide molecules per plasmid. This repquine, added to assist endocytic exit, unexpectedly resented approximately a 6:1 ratio of positive to negative reduced transfection efficiency in two of the cell lines charges. Peptide 5 was shown to have a higher transfectested, COS-7 and ECV304. However, transfection tion efficiency under most conditions, possibly because of efficiency increased at higher chloroquine concentrations more efficient stabilisation of cyclisation by two cysteineand exceeded that in the absence of chloroquine in the cysteine bonds.

show abstract

Identification of the Integrin VLA-2 as a Receptor for Echovirus 1

Cited by 288 publications

References 31 publications

Mapping the Collagen-binding Site in the I Domain of the Glycoprotein Ia/IIa (Integrin α2β1)

Mapping the Collagen-binding Site in the I Domain of the Glycoprotein Ia/IIa (Integrin α2β1)

Clustering Induces a Lateral Redistribution of α2β1 Integrin from Membrane Rafts to Caveolae and Subsequent Protein Kinase C-dependent Internalization

Integrin-mediated transfection with peptides containing arginine-glycine-aspartic acid domains

Contact Info

Product

Resources

About