Vascular endothelial cells assemble adhesive intercellular junctions comprising a unique cadherin, VE-cadherin, which is coupled to the actin cytoskeleton through cytoplasmic interactions with plakoglobin, beta-catenin and alpha -catenin. However, the potential linkage between VE-cadherin and the vimentin intermediate filament cytoskeleton is not well characterized. Recent evidence indicates that lymphatic and vascular endothelial cells express desmoplakin, a cytoplasmic desmosomal protein that attaches intermediate filaments to the plasma membrane in epithelial cells. In the present study, desmoplakin was localized to intercellular junctions in human dermal microvascular endothelial cells. To determine if VE-cadherin could associate with desmoplakin, VE-cadherin, plakoglobin, and a desmoplakin amino-terminal polypeptide (DP-NTP) were co-expressed in L-cell fibroblasts. In the presence of VE-cadherin, both plakoglobin and DP-NTP were recruited to cell-cell borders. Interestingly, beta-catenin could not substitute for plakoglobin in the recruitment of DP-NTP to cell borders, and DP-NTP bound to plakoglobin but not beta-catenin in the yeast two-hybrid system. In addition, DP-NTP colocalized at cell-cell borders with alpha-catenin in the L-cell lines, and endogenous desmoplakin and alpha-catenin colocalized in cultured dermal microvascular endothelial cells. This is in striking contrast to epithelial cells, where desmoplakin and alpha -+catenin are restricted to desmosomes and adherens junctions, respectively. These results suggest that endothelial cells assemble unique junctional complexes that couple VE-cadherin to both the actin and intermediate filament cytoskeleton.

REFERENCES

Aberle
H.
,
Butz
S.
,
Stappert
J.
,
Weissig
H.
,
Kemler
R.
,
Hoschuetzky
H.
(
1994
).
Assembly of the cadherin-catenin complex in vitro with recombinant proteins.
J. Cell Sci
107
,
3655
3663
Albelda
S. M.
,
Muller
W. A.
,
Buck
C. A.
,
Newman
P. J.
(
1991
).
Molecular and cellular properties of PECAM-1 (endoCAM/CD31): A novel vascular cell-cell adhesion molecule.
J. Cell Biol
114
,
1059
1068
Ali
J.
,
Liao
F.
,
Martens
E.
,
Muller
W. A.
(
1997
).
Vascular endothelial cadherin (VE-cadherin): cloning and role in endothelial cell-cell adhesion.
Microcirculation
4
,
267
277
Angst
B. D.
,
Nilles
L. A.
,
Green
K. J.
(
1990
).
Desmoplakin II expression is not restricted to stratified epithelia.
J. Cell Sci
97
,
247
257
Bach
T. L.
,
Barsigian
C.
,
Chalupowicz
D. G.
,
Busler
D.
,
Yaen
C. H.
,
Grant
D. S.
,
Martinez
J.
(
1998
).
VE-cadherin mediates endothelial cell capillary tube formation in fibrin and collagen gels.
Exp. Cell Res
238
,
324
334
Bai
C.
,
Elledge
S. J.
(
1995
).
Cloning using the two-hybrid system.
Meth. Enzymol
273
,
331
347
Barth
A. I.
,
Nathke
I. S.
,
Nelson
W. J.
(
1997
).
Cadherins, catenins and APC protein: interplay between cytoskeletal complexes and signaling pathways.
Curr. Opin. Cell Biol
9
,
683
690
Bornslaeger
E. A.
,
Corcoran
C. M.
,
Stappenbeck
T. S.
,
Green
K. J.
(
1996
).
Breaking the connection: Displacement of the desmosomal plaqueprotein desmoplakin from cell-cell interfaces disrupts anchorage of intermediate filament bundles and alters intercellular junction assembly.
J. Cell Biol
134
,
985
1001
Breviario
F.
,
Caveda
L.
,
Corada
M.
,
Martin-Padura
I.
,
Navarro
P.
,
Golay
J.
,
Introna
M.
,
Gulino
D.
,
Lampugnani
M. G.
,
Dejana
E.
(
1995
).
Functional properties of human vascular endothelial cadherin (7B4/Cadherin-5), an endothelium-specific cadherin.
Arterioscler. Thromb. Vasc. Biol
15
,
1229
1239
Cowin
P.
,
Kapprell
H.
,
Franke
W. W.
,
Tamkun
J. W.
,
Hynes
R. O.
(
1986
).
Plakoglobin: A protein common to different kinds of intercellular adhering junctions.
Cell
46
,
1063
1073
Cowin
P.
,
Burke
B.
(
1996
).
Cytoskeleton-membrane interactions.
Curr. Opin. Cell Biol
8
,
56
65
Dejana
E.
,
Corada
M.
,
Lampugnani
M. G.
(
1995
).
Endothelial cell-cell junctions.
FASEB J
9
,
910
918
Dejana
E.
(
1996
).
Endothelial adherens junctions: implications in the control of vascular permeability and angiogenesis.
J. Clin. Invest
98
,
1949
1953
Fuchs
E.
(
1994
).
Intermediate filaments and disease: mutations that cripple cell strength.
J. Cell Biol
125
,
511
516
Fuchs
E.
,
Cleveland
D. W.
(
1998
).
A structural scaffolding of intermediate filaments in health and disease.
Science
279
,
514
519
Garrod
D.
,
Chidgey
M.
,
North
A.
(
1996
).
Desmosomes: differentiation, development dynamics and disease.
Curr. Opin. Cell Biol
8
,
670
678
Green
K. J.
,
Jones
J. C. R.
(
1996
).
Desmosomes and hemidesmosomes: structure and function of molecular components.
FASEB J
10
,
871
881
Green
K. J.
,
Bornslaeger
E. A.
,
Kowalczyk
A. P.
,
Palka
H. L.
,
Norvell
S. M.
(
1997
).
Specificity of desmosomal plaque protein interactions with intermediate filaments: keeping adhesive junctions segregated.
J. Gen. Physiol
52
,
123
139
Harper
J. W.
,
Adami
G.
,
Wei
N.
,
Keyomarsi
K.
,
Elledge
S. J.
(
1993
).
The p21 Cdk-interacting protein Cip1 is a potent inhibitor of G1 cyclin-dependent kinases.
Cell
75
,
805
816
Herrenknecht
K.
,
Ozawa
M.
,
Eckerskorn
C.
,
Lottspeich
F.
,
Lenter
M.
,
Kemler
R.
(
1991
).
The uvomorulin-anchorage protein alpha catenin is a vinculin homologue.
Proc. Nat. Acad. Sci. USA
88
,
9156
9160
Hinck
L.
,
Nelson
W. J.
,
Papkoff
J.
(
1994
).
Wnt-1 modulates cell-cell adhesion in mammalian cells by stabilizing-catenin binding to the cell adhesion protein cadherin.
J. Cell Biol
124
,
729
741
Huber
O.
,
Krohn
M.
,
Kemler
R.
(
1997
).
A specific domain in-catenin medites binding to -catenin or plakoglobin.
J. Cell Sci
110
,
1759
1765
Jou
T.-S.
,
Stewart
D. B.
,
Stappert
J.
,
Nelson
W. J.
,
Marrs
J. A.
(
1995
).
Genetic and biochemical dissection of protein linkages in the cadherin-catenin complex.
Proc. Nat. Acad. Sci USA
92
,
5067
5071
Klymkowsky
M. W.
,
Parr
B.
(
1995
).
The body language of cells: The intimate connection between cell adhesion and behavior.
Cell
83
,
5
8
Knudsen
K. A.
,
Wheelock
M. J.
(
1992
).
Plakoglobin, or an 83-kD homologue distinct from-catenin, interacts with E-cadherin and N-cadherin.
J. Cell Biol
118
,
671
679
Knudsen
K. A.
,
Soler
A. P.
,
Johnson
K. R.
,
Wheelock
M. J.
(
1995
).
Interaction of-actinin with the cadherin/catenin cell-cell adhesion complex via -catenin.
J. Cell Biol
130
,
67
77
Kouklis
P. D.
,
Hutton
E.
,
Fuchs
E.
(
1994
).
Making a connection: Direct binding between keratin intermediate filaments and desmosomal proteins.
J. Cell Biol
127
,
1049
1060
Kowalczyk
A. P.
,
Palka
H. L.
,
Luu
H. H.
,
Nilles
L. A.
,
Anderson
J. E.
,
Wheelock
M. J.
,
Green
K. J.
(
1994
).
Posttranslational regulation of plakoglobin expression: Influence of the desmosomal cadherins on plakoglobin metabolic stability.
J. Biol. Chem
269
,
31214
31223
Kowalczyk
A. P.
,
Borgwardt
J. E.
,
Green
K. J.
(
1996
).
Analysis of desmosomal cadherin adhesive function and stoichiometry of the desmosomal cadherin:plakoglobin complex.
J. Invest. Dermatol
107
,
293
300
Kowalczyk
A. P.
,
Bornslaeger
E. A.
,
Borgwardt
J. E.
,
Palka
H. L.
,
Dhaliwal
A. S.
,
Corcoran
C. M.
,
Denning
M. F.
,
Green
K. J.
(
1997
).
The amino-terminal domain of desmoplakin binds to plakoglobin and clusters desmosomal cadherin-plakoglobin complexes.
J. Cell Biol
139
,
773
784
Lampugnani
M. G.
,
Corada
M.
,
Caveda
L.
,
Breviario
F.
,
Ayalon
O.
,
Geier
B.
,
Dejana
E.
(
1995
).
The molecular organization of endothelial cell-cell junctions: Differential association of plakoglobin,-catenin, and -catenin with vascular endothelial cadherin (VE-cadherin).
J. Cell Biol
129
,
203
217
Lampugnani
M. G.
,
Dejana
E.
(
1997
).
Interendothelial junctions: structure, signaling and functional roles.
Curr. Opin. Cell Biol
9
,
674
682
Lum
H.
,
Malik
A. B.
(
1996
).
Mechanisms of increased endothelial permeability.
Can. J. Physiol. Pharmacol
74
,
787
800
Mathur
M.
,
Goodwin
L.
,
Cowin
P.
(
1994
).
Interactions of the cytoplasmic domain of the desmosomal cadherin Dsg1 with plakoglobin.
J. Biol. Chem
269
,
14075
14080
Matsumura
T.
,
Wolff
K.
,
Petzelbauer
P.
(
1997
).
Endothelial cell tube formation depends on cadherin-5 and CD31 interactions with filamentous actin.
J. Immunol
158
,
3408
3416
Meng
J.-J.
,
Bornslaeger
E. A.
,
Green
K. J.
,
Steinert
P. M.
,
Ip
W.
(
1997
).
Two-hybrid analysis reveals fundamental differences in direct interactions between desmoplakin and cell type-specific intermediate filaments.
J. Biol. Chem
272
,
21495
21503
Minnear
F. L.
,
DeMichele
M. A.
,
Moon
D. G.
,
Reider
C. L.
,
Fenton
J. W.
II
(
1989
).
Isoproterenol reduces thrombin-induced pulmonary endothelial permeability in vitro.
Am. J. Physiol
257
,
1613
–.
Nagafuchi
A.
,
Takeichi
M.
,
Tsukita
S.
(
1991
).
The 102kd cadherin-associated protein: similarity to vinculin and postranscriptional regulation of expression.
Cell
65
,
849
857
Navarro
P.
,
Caveda
L.
,
Breviario
F.
,
Mandoteanu
I.
,
Lampugnani
M. G.
,
Dejana
E.
(
1995
).
Catenin-dependent and-independent functions of vascular endothelial cadherin.
J. Biol. Chem
52
,
30965
30972
Navarro
P.
,
Ruco
L.
,
Dejana
E.
(
1998
).
Differential localization of VE-and N-cadherins in human endothelial cells: VE-cadherin competes with N-cadherin for junctional localization.
J. Cell Biol
140
,
1475
1484
Newman
P. J.
,
Berndt
M. C.
,
Gorski
J.
,
White
G. C.
II
,
Lyman
S.
,
Paddock
C.
,
Muller
W. A.
(
1990
).
PECAM-1 (CD-31) cloning and relation to adhesion molecules of the immunoglobulin gene superfamily.
Science
247
,
1219
1222
Newman
P. J.
(
1997
).
The biology of PECAM-1.
J. Clin. Invest
99
,
3
8
Nieset
J. E.
,
Redfield
A. R.
,
Jin
F.
,
Knudsen
K. A.
,
Johnson
K. R.
,
Wheelock
M. J.
(
1997
).
Characterization of the interactions of-catenin with -actinin and-catenin/plakoglobin.
J. Cell Sci
110
,
1013
1022
Norvell
S. M.
,
Green
K. J.
(
1998
).
Contributions of extracellular and intracellular domains of full length and chimeric cadherin molecules to junction assembly in epithelial cells.
J. Cell Sci
111
,
1305
1318
Obama
H.
,
Ozawa
M.
(
1997
).
Identification of the domain of-catenin involved in its association with -catenin and plakoglobin (-catenin).
J. Biol. Chem
272
,
11017
11020
Ozawa
M.
,
Baribault
H.
,
Kemler
R.
(
1989
).
The cytoplasmic domain of the cell adhesion molecule uvomorulin associates with three independent proteins structurally related in different species.
EMBO J
8
,
1711
1717
Ozawa
M.
,
Kemler
R.
(
1992
).
Molecular organization of the uvomorulin-catenin complex.
J. Cell Biol
116
,
989
996
Palka
H. L.
,
Green
K. J.
(
1997
).
Roles of plakoglobin end domains in desmosome assembly.
J. Cell Sci
110
,
2359
2371
Plott
R. T.
,
Amagai
M.
,
Udey
M. C.
,
Stanley
J. R.
(
1994
).
Pemphigusvulgaris antigen lacks biochemical properties characteristic of classical cadherins.
J. Invest. Dermatol
103
,
168
172
Rimm
D. L.
,
Koslov
E. R.
,
Kebriaei
P.
,
Cianci
C. D.
,
Morrow
J. S.
(
1995
).
(E)-catenin is an actin-binding and-bundling protein mediating the attachment of F-actin to the membrane adhesion complex.
Proc. Nat. Acad. Sci USA
92
,
8813
8817
Roh
J.-Y.
,
Stanley
J. R.
(
1995
).
Plakoglobin binding by human Dsg3 (pemphigus vulgaris antigen) in keratinocytes requires the cadherin-like intracytoplasmic segment.
J. Invest. Dermatol
104
,
720
724
Savion
N.
,
Vlodavsky
I.
,
Greenburg
G.
,
Gospodarowicz
D.
(
1982
).
Synthesis and distribution of cytoskeletal elements in endothelial cells as a function of cell growth and organization.
J. Cell Physiol
110
,
129
141
Schmelz
M.
,
Franke
W. W.
(
1993
).
Complexus adhaerentes, a new group of desmoplakin-containing junctions in endothelial cells: the syndesmos connecting retothelial cells of lymph nodes.
Eur. J. Cell Biol
61
,
274
289
Schmelz
M.
,
Moll
R.
,
Kuhn
C.
,
Franke
W. W.
(
1994
).
Complexus adhaerentes, a new group of desmoplakin-containing junctions in endothelial cells: II. Different types of lymphatic vessels.
Differentiation
57
,
97
117
Schnittler
H.-J.
,
Puschel
B.
,
Drenckhahn
D.
(
1997
).
Role of cadherins and plakoglobin in interendothelial adhesion under resting conditions and shear stress.
Am. J. Physiol
273
,
2396
–.
Smith
E. A.
,
Fuchs
E.
(
1998
).
Defining the interactions between intermediate filaments and desmosomes.
J. Cell Biol
141
,
1229
1241
Stappenbeck
T. S.
,
Green
K. J.
(
1992
).
The desmoplakin carboxyl terminus coaligns with and specifically disrupts intermediate filament networks when expressed in cultured cells.
J. Cell Biol
116
,
1197
1209
Stappenbeck
T. S.
,
Bornslaeger
E. A.
,
Corcoran
C. M.
,
Luu
H. H.
,
Virata
M. L. A.
,
Green
K. J.
(
1993
).
Functional analysis of desmoplakin domains: Specification of the interaction with keratin versus vimentin intermediate filament networks.
J. Cell Biol
123
,
691
705
Stappenbeck
T. S.
,
Lamb
J. A.
,
Corcoran
C. M.
,
Green
K. J.
(
1994
).
Phosphorylation of the desmoplakin COOH terminus negatively regulates its interaction with keratin intermediate filament networks.
J. Biol. Chem
269
,
29351
29354
Stasek
J. E.
Jr.
,
Patterson
C. E.
,
Garcia
J. G.
(
1992
).
Protein kinase C phosphorylates caldesmon77 and vimentin and enhances albumin permeability across cultured bovine pulmonary artery endothelial cell monolayers.
J. Cell Physiol
153
,
62
75
Steinert
P. M.
,
Bale
S. J.
(
1993
).
Genetic skin diseases caused by mutations in keratin intermediate filaments.
Trends Genet
9
,
280
284
Tanihara
H.
,
Kido
M.
,
Obata
S.
,
Heimark
R. L.
,
Davidson
M.
,
St John
T.
,
Suzuki
S.
(
1994
).
Characterization of cadherin-4 and cadherin-5 reveals new aspects of cadherins.
J. Cell Sci
107
,
1694
1704
Telo
P.
,
Breviario
F.
,
Huber
P.
,
Panzeri
C.
,
Dejana
E.
(
1998
).
Identification of a novel cadherin (vascular endothelial cadherin-2) located at intercellular junctions in endothelial cells.
J. Biol. Chem
273
,
17565
17572
Valiron
O.
,
Chevier
V.
,
Usson
Y.
,
Breviario
F.
,
Job
D.
,
Dejana
E.
(
1996
).
Desmoplakin expression and organization at human umbilical vein endothelial cell-cell junctions.
J. Cell Sci
109
,
2141
2149
van Hinsbergh
V. M. M.
(
1997
).
Endothelial permeability for macromolecules.
Arterioscler. Thromb. Vasc. Biol
17
,
1018
1023
Vittet
D.
,
Buchou
T.
,
Schweitzer
A.
,
Dejana
E.
,
Huber
P.
(
1997
).
Targetted null-mutation in the vascular endothelial-cadherin gene impairs the organization of vascular-like structures in embryoid bodies.
Proc. Nat. Acad. Sci. USA
94
,
6273
6278
This content is only available via PDF.