The cytoplasmic domain of the transmembrane protein M-cadherin is involved in anchoring cytoskeletal elements to the plasma membrane at cell-cell contact sites. Several members of the armadillo repeat protein family mediate this linkage. We show here that ARVCF, a member of the p120 (ctn) subfamily, is a ligand for the cytoplasmic domain of M-cadherin, and characterize the regions involved in this interaction in detail. Complex formation in an in vivo environment was demonstrated in (1) yeast two-hybrid screens, using a cDNA library from differentiating skeletal muscle and part of the cytoplasmic M-cadherin tail as a bait, and (2) mammalian cells, using a novel experimental system, the MOM recruitment assay. Immunoprecipitation and in vitro binding assays confirmed this interaction. Ectopically expressed EGFP-ARVCF-C11, an N-terminal truncated fragment, targets to junctional structures in epithelial MCF7 cells and cardiomyocytes, where it colocalizes with the respective cadherins, beta-catenin and p120 (ctn). Hence, the N terminus of ARVCF is not required for junctional localization. In contrast, deletion of the four N-terminal armadillo repeats abolishes this ability in cardiomyocytes. Detailed mutational analysis revealed the armadillo repeat region of ARVCF as sufficient and necessary for interaction with the 55 membrane-proximal amino acids of the M-cadherin tail.

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
Allport
J. R.
,
Muller
W. A.
,
Luscinskas
F. W.
(
2000
).
Monocytes induce reversible focal changes in vascular endothelial cadherin complex during transendothelial migration under flow.
J. Cell Biol
148
,
203
216
Anastasiadis
P. Z.
,
Reynolds
A. B.
(
2000
).
The p120 catenin family: complex roles in adhesion, signaling and cancer.
J. Cell Sci
113
,
1319
1334
Aono
S.
,
Nakagawa
S.
,
Reynolds
A. B.
,
Takeichi
M.
(
1999
).
p120(ctn) acts as an inhibitory regulator of cadherin function in colon carcinoma cells.
J. Cell Biol
145
,
551
562
Behrens
J.
,
von Kries
J. P.
,
Kuhl
M.
,
Bruhn
L.
,
Wedlich
D.
,
Grosschedl
R.
,
Birchmeier
W.
(
1996
).
Functional interaction of-catenin with the transcription factor LEF-1.
Nature
382
,
638
642
Brieher
W. M.
,
Yap
A. S.
,
Gumbiner
B. M.
(
1996
).
Lateral dimerization is required for the homophilic binding activity of C-cadherin.
J. Cell Biol
135
,
487
496
Butz
S.
,
Kemler
R.
(
1994
).
Distinct cadherin-catenin complexes in Ca(2+)-dependent cell-cell adhesion.
FEBS Lett
355
,
195
200
Daniel
J. M.
,
Reynolds
A. B.
(
1999
).
The catenin p120(ctn) interacts with kaiso, a novel BTB/POZ domain zinc finger transcription factor.
Mol. Cell Biol
19
,
3614
3623
Donalies
M.
,
Cramer
M.
,
Ringwald
M.
,
Starzinski-Powitz
A.
(
1991
).
Expression of M-cadherin, a member of the cadherin multigene family, correlates with differentiation of skeletal muscle cells.
Proc. Natl. Acad. Sci. USA
88
,
8024
8028
Eger
A.
,
Stockinger
A.
,
Schaffhauser
B.
,
Berg
H.
,
Foisner
R.
(
2000
).
Epithelial mesenchymal transition by c-Fos estrogen receptor activation involves nuclear translocation of beta-catenin and upregulation of beta-catenin / lymphoid enhancer binding factor-1 transcriptional activity.
J. Cell Biol
148
,
173
188
Eppenberger-Ebehardt
M.
,
Flamme
I.
,
Kurer
V.
,
Eppenberger
H. M.
(
1990
).
Reexpression of alpha-smooth muscle actin isoform in cultured adult rat cardiomyocytes.
Dev. Biol
139
,
269
278
Eppenberger
H. M.
,
Zuppinger
C.
(
1999
).
In vitro reestablishment of cell-cell contacts in adult rat cardiomyocytes. Functional role of transmembrane components in the formation of new intercalated disc-like cell contacts.
FASEBJ
13
,
83
89
Funke
B.
,
Saint-Jore
B.
,
Puech
A.
,
Sirotkin
H.
,
Edelmann
L.
,
Carlson
C.
,
Raft
S.
,
Pandita
R. K.
,
Kucherpalati
R.
,
Skoultchi
A.
,
Morrow
B. E.
(
1997
).
Characterization and mutation analysis of goosecoid-like (GSCL), a homeodomain-containing gene that maps to the critical region for VCFS/DGS on 22q11.
Genomics
46
,
364
372
Funke
B.
,
Puech
A.
,
Saint-Jore
B.
,
Pandita
R.
,
Skoultchi
A.
,
Morrow
B.
(
1998
).
Isolation and characterization of a human gene containing a nuclear localization signal from the critical region for velo-cardio-facial syndrome on 22q11.
Genomics
53
,
146
154
Finnemann
S.
,
Mitrik
I.
,
Hess
M.
,
Otto
G.
,
Wedlich
D.
(
1997
).
Uncoupling of XB/U-cadherin-catenin complex formation from its function in cell cell adhesion.
J. Biol. Chem
272
,
11856
11862
Gluzman
Y.
(
1981
).
SV40-transformed simian cells support the replication of early SV40 mutants.
Cell
23
,
175
182
Grove
B. K.
,
Kurer
V.
,
Lehner
C.
,
Doetschman
T. C.
,
Perriard
J. C.
,
Eppenberger
H. M.
(
1984
).
A new 185,000-dalton skeletal muscle protein detected by monoclonal antibodies.
J. Cell Biol
98
,
518
524
Gumbiner
B. M.
(
1996
).
Cell adhesion: The molecular basis of tissue architecture and morphogenisis.
Cell
84
,
345
357
Hatzfeld
M.
,
Nachtsheim
C.
(
1996
).
Cloningand characterization of a new armadillo family member, p0071, associated with the junctional plaque:evidence for a subfamily of closely related proteins.
J. Cell Sci
109
,
2767
2778
Hatzfeld
M.
(
1999
).
The armadillo family of structural proteins.
Int. Rev. Cytol
186
,
179
224
Hazan
R. B.
,
Norton
L.
(
1998
).
The epidermal growth factor receptor modulates the interaction of E-cadherin with the actin cytoskeleton.
J. Biol. Chem
273
,
9078
9084
Heisterkamp
N.
,
Mulder
M. P.
,
Langeveld
A.
,
ten Hoeve
J.
,
Wang
Z.
,
Roe
B. A.
,
Groffen
J.
(
1995
).
Localization of the human mitochondrial citrate transporter protein gene to chromosome 22q11 in the DiGeorge syndrome critical region.
Genomics
29
,
451
456
Hertig
C. M.
,
Butz
S.
,
Koch
S.
,
Eppenberger-Ebehardt
M.
,
Kemler
R.
,
Eppenberger
H. M.
(
1996
).
N-cadherin in adult rat cardiomyocytes in culture.
J. Cell Sci
109
,
11
20
Hill
K.
,
Model
K.
,
Ryan
M. T.
,
Dietmeier
K.
,
Martin
F.
,
Wagner
R.
,
Pfanner
N.
(
1998
).
Tom40 forms the hydrophilic channel of the mitochondrial import pore for preproteins.
Nature
395
,
516
521
Hinck
L.
,
Näthke
I. S.
,
Papkoff
J.
,
Nelson
W. J.
(
1994
).
Dynamics of cadherin-catenin complex formation: novel protein interactions and pathways of complex assembly.
J. Cell Biol
125
,
1327
1340
Hirano
S.
,
Nose
A.
,
Hatta
K.
,
Kawakami
A.
,
Takeichi
M.
(
1987
).
Calcium-dependent cell-cell adhesion molecules (cadherins): subclass specifities and possible involvement of actin bundles.
J. Cell Biol
105
,
2501
2510
Hirano
S.
,
Kimoto
N.
,
Shimoyama
Y.
,
Hirohashi
S.
,
Takeichi
M.
(
1992
).
Identification of a neural-catenin as a key regulator of cadherin function and multicellular organization.
Cell
70
,
293
301
Huber
O.
,
Bierkamp
C.
,
Kemler
R.
(
1996
).
Cadherins and catenins in development.
Curr. Opin. Cell Biol
8
,
685
691
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
Kuch
C.
,
Winnekendonk
D.
,
Butz
S.
,
Unvericht
U.
,
Kemler
R.
,
Starzinski-Powitz
A.
(
1997
).
M-cadherin-mediated cell adhesion and complex formation with the catenins in myogenic mouse cells.
Exp. Cell Res
232
,
331
338
Kudo
N.
,
Matsumori
N.
,
Taoka
H.
,
Fujiwara
D.
,
Schreiner
E. P.
,
Wolff
B.
,
Yoshida
M.
,
Horinouchi
S.
(
1999
).
Leptomycin B inactivates CRM/exportin 1 by covalent modification at a cysteine residue in the central conserved region.
Proc. Natl. Acad. Sci. USA
96
,
9112
9117
Lampugnani
M. G.
,
Corada
M.
,
Andriopoulou
P.
,
Esser
S.
,
Risau
W.
,
Dejana
E.
(
1997
).
Cell confluence regulates tyrosine phosphorylation of adherens junction components in endothelial cells.
J. Cell Sci
110
,
2065
2077
Lindsay
E. A.
,
Botta
A.
,
Jurecic
V.
,
Carattini-Rivera
S.
,
Cheah
Y. C.
,
Rosenblatt
H. M.
,
Bradley
A.
,
Baldini
A.
(
1999
).
Congenital heart disease in mice for the DiGeorge syndrome region.
Nature
401
,
379
383
Lu
Q.
,
Paredes
M.
,
Medina
M.
,
Zhou
J.
,
Cavallo
R.
,
Peifer
M.
,
Orecchio
L.
,
Kosik
K. S.
(
1999
).
Delta-catenin, an adhesive junction-associated protein which promotes cell scattering.
J. Cell Biol
144
,
519
532
Mariner
D. J.
,
Wang
J.
,
Reynolds
A. B.
(
2000
).
ARVCF localizes to the nucleus and adherens junction and is mutually exclusive with p120ctn in E-cadherin complexes.
J. Cell Sci
113
,
1481
1490
McBride
H. M.
,
Millar
D. G.
,
Li
J. M.
,
Shore
G. C.
(
1992
).
A signal-anchor sequence selective for the mitochondrial outer membrane.
J. Cell Biol
119
,
1451
1457
Messerli
G. M.
,
van der Voort
H. T. M.
,
Rungger-Brindle
E.
,
Perriard
G.-C.
) (
1993
).
Three-dimensional visualization of multi-channel voxel data.
Cytometry
14
,
725
735
Molenaar
M.
,
van de Wetering
M.
,
Oosterwegel
M.
,
Peterson-Maduro
J.
,
Godsave
S.
,
Korinek
V.
,
Roose
J.
,
Destree
O.
,
Clevers
H.
(
1996
).
XTcf-3 transcription factor mediates-catenin-induced axis formation in Xenopus embryos.
Cell
86
,
391
399
Muller
T.
,
Choidas
A.
,
Reichmann
E.
,
Ulrich
A.
(
1999
).
Phosphorylation and free pool of-catenin are regulated by tyrosine kinases and tyrosine phosphatases during epithelial cell migration.
J. Biol. Chem
274
,
10173
10183
Näthke
I.
,
Hinck
S.
,
Swedlow
J. R.
,
Papkoff
J.
,
Nelson
J. W.
(
1994
).
Defining interactions and distributions of cadherin and catenin complexes in polarized epithelial cells.
J. Cell Biol
125
,
1341
1352
Nagar
B.
,
Overduin
M.
,
Ikura
M.
,
Rini
J. M.
(
1996
).
Structural basis of calcium-induced E-cadherin rigidification and dimerization.
Nature
380
,
360
364
Nieset
J. E.
,
Redfield
A. R.
,
Jin
F.
,
Knudsen
K.
,
Johnson
K. R.
,
Wheelock
M. J.
(
1997
).
Characterization of the interactions of-catenin with -actinin and-catenin/plakoglobin.
J. Cell Sci
110
,
1013
1022
Ohkubo
T.
,
Ozawa
M.
(
1999
).
P120(ctn) binds to the membrane-proximal region of the E-cadherin cytoplasmic domain and is involved in modulation of adhesion activity.
J. Biol. Chem
274
,
21409
21415
Orsulic
M.
,
Huber
O.
,
Aberle
H.
,
Arnold
S.
,
Kemler
R.
(
1999
).
E-cadherion binding prevents-catenin nuclear localization and -catenin/LEF-1 mediated transactivation.
J. Cell Sci
112
,
1237
1245
Ozawa
M.
,
Baribault
H.
,
Kemler
R.
(
1989
).
The cytoplasmatic 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.
(
1998
).
The membrane-proximal region of the E-cadherin cytoplasmic domain prevents dimerization and negatively regulates adhesion activity.
J. Cell Biol
142
,
1605
1613
Paffenholz
R.
,
Franke
W.
(
1997
).
Identification and loclization of a neurally expressed member of the plakoglobin/armadillo multigene family.
Differentiation
61
,
293
304
Pertz
O.
,
Bozic
D.
,
Koch
A. W.
,
Fauser
C.
,
Brancaccio
A.
,
Engel
J.
(
1999
).
A new crystal structure, Ca2+ dependence and mutational analysis reveal molecular details of E-cadherin homoassociation.
EMBO J
18
,
1738
1747
Pfanner
N.
,
Meijer
M.
(
1997
).
The Tom and Tim machine.
Curr. Biol
7
,
100
103
Provost
E.
,
Rimm
D. L.
(
1999
).
Controversies at the cytoplasmic face of the cadherin-based adhesion complex.
Curr. Opin. Cell Biol
11
,
567
572
Puech
A.
,
Saint-Jore
B.
,
Funke
B.
,
Gilbert
D. J.
,
Sirotkin
H.
,
Copeland
N. G.
,
Jenkins
N. A.
,
Kucherlapati
R.
,
Morrow
B.
,
Skoultchi
A.
(
1997
).
Comparative mapping of the human 22q11 chromosomal region and the orthologous region in mice reveals complex changes in gene organization.
Proc. Natl. Acad. Sci. USA
94
,
14608
14613
Reynolds
A. B.
,
Daniel
J.
,
McCrea
P. D.
,
Wheelock
M. J.
,
Wu
J.
,
Zhang
Z.
(
1994
).
Identification of a new catenin: the tyrosine kinase substrate p120cas associates with E-cadherin complexes.
Mol. Cell. Biol
14
,
8333
8342
Reynolds
A. B.
,
Daniel
J.
,
Mo
Y. Y.
,
Wu
J.
,
Zhang
Z.
(
1996
).
The novel catenin p120cas binds classical cadherins and induces an unusual morphological phenotype in NIH3T3 fibroblasts.
Exp. Cell Res
225
,
328
337
Riese
J.
,
Yu
X.
,
Munnerlyn
A.
,
Eresh
S.
,
Hsu
S. C.
,
Grosschedl
R.
,
Bienz
M.
(
1997
).
LEF-1, a nuclear factor coordinating signaling inputs from wingless and decapentaple.
Cell
88
,
777
787
Rimm
D. l.
,
Koslov
E. R.
,
Kebriaei
P.
,
Cianci
C. D.
,
Morrow
J. S.
(
1995
).
Alpha 1(E)-catenin is an actin-binding and-bundling protein mediating the attachment of F-actin to the membrane adhesion complex.
Proc. Natl. Acad. Sci. USA
92
,
8813
8817
Ringwald
M.
,
Schuh
R.
,
Vestweber
D.
,
Eistetter
H.
,
Lottspeich
F.
,
Engel
J.
,
Dölz
R.
,
Jähnig
F.
,
Epplen
J.
,
Mayer
S.
,
Muller
C.
,
Kemler
R.
(
1987
).
The structure of the cell adhesion molecule uvomorulin. Insights into the molecular mechanism of Ca2+ dependent cell adhesion.
EMBO J
6
,
3647
3653
Rose
O.
,
Rohwedel
J.
,
Reinhardt
S.
,
Bachmann
S.
,
Cramer
M.
,
Rotter
M.
,
Wobus
M.
,
Starzinski-Powitz
A.
(
1994
).
Expression of M-cadherin protein in myogenic cells during prenatal mouse development and differentiation of embryonic stem cells in culture.
Dev. Dyn
201
,
245
259
Rudiger
M.
,
Jockusch
B. M.
,
Rothkegel
M.
(
1997
).
A novel epitope-antibody combination for the detection of protein expression in prokaryotic and eukaryotic cells.
Biotechniques
23
,
96
97
Schatz
G.
(
1998
).
The doors to organelles.
Nature
395
,
439
440
Shapiro
L.
,
Fannon
A. M.
,
Kwong
P. D.
,
Thompson
A.
,
Lehmann
M. S.
,
Grubel
G.
,
Legrand
I.
,
Als-Nielsen
J.
,
Colman
D. R.
,
Hendrickson
W. A.
(
1995
).
Structural basis of cell-cell adhesion by cadherins.
Nature
374
,
327
336
Sirotkin
H.
,
Morrow
B.
,
DasGupta
R.
,
Goldberg
R.
,
Patanjali
S. R.
,
Shi
G.
,
Cannizzaro
L.
,
Shprintzen
R.
,
Weissman
S. M.
,
Kucherlapati
R.
(
1996
).
Isolation of a new clathrin heavy chain gene with muscle-specific expression from the region commonly deleted in velo-cardio-facial syndrome.
Hum. Mol. Genet
5
,
617
624
Sirotkin
H.
,
O'Donnell
H.
,
DasGupta
R.
,
Halford
S.
,
St Jore
B.
,
Puech
A.
,
Parimoo
S.
,
Morrow
B.
,
Skoultchi
A.
,
Weissman
S. M.
,
Scambler
P.
,
Kucherlapati
R.
(
1997
).
Identification of a new human catenin gene family member (ARVCF) from the region deleted in velo-cardio-facial syndrome.
Genomics
41
,
75
83
Sirotkin
H.
,
Morrow
B.
,
Saint-Jore
B.
,
Puech
A.
,
Das Gupta
R.
,
Patanjali
S. R.
,
Skoultchi
A.
,
Weissman
S. M.
,
Kucherlapati
R.
(
1997
).
Identification, characterization, and precise mapping of a human gene encoding a novel membrane-spanning protein from the 22q11region deleted in velo-cardio-facial syndrome.
Genomics
42
,
245
251
Staddon
J. M.
,
Smales
C.
,
Schulze
C.
,
Esch
F. S.
,
Rubin
L. L.
(
1995
).
p120, a p120-related protein (p100), and the cadherin/catenin complex.
J. Cell Biol
130
,
369
381
Steinberg
M. S.
,
McNutt
P. M.
(
1999
).
Cadherins and their connections: Adhesion junctions have broader functions.
Curr. Opin. Cell Biol
11
,
545
560
Thoreson
M. A.
,
Annastasiadis
P. Z.
,
Daniel
J. M.
,
Ireton
R. C.
,
Wheelock
M. J.
,
Johnson
K. R.
,
Hummingbird
D. K.
,
Reynolds
A. B.
(
2000
).
Selective uncoupling of p120 (ctn) from E-cadherin disrupts strong adhesion.
J. Cell Biol
148
,
189
202
van de Wetering
M.
,
Cavallo
R.
,
Dooijes
D.
,
van Beest
M.
,
van Es
J.
,
Loureiro
J.
,
Ypma
A.
,
Hursh
D.
,
Jones
T.
,
Bejsovec
A.
,
Peifer
M.
,
Mortin
M.
,
Clevers
H.
(
1997
).
Armadillo coactivates transcription driven by the product of the Drosophila segment polarity gene dTCF.
Cell
88
,
789
799
van Hengel
J.
,
Vanhoenacker
P.
,
Staes
K.
,
van Roy
F.
(
1999
).
Nuclear localization of the p120 ctn Armadillo-like catenin is counteracted by a nuclear export signal and by E-cadherin expression.
Proc. Natl. Acad. Sci. USA
96
,
7980
7985
Vleminckx
K.
,
Kemler
R.
(
1999
).
Cadherins and tissue formation: integrating adhesion and signaling.
BioEssays
21
,
211
220
Watabe-Uchida
M.
,
Uchida
N.
,
Imamura
Y.
,
Nagafuchi
A.
,
Fujimoto
K.
,
Uemura
T.
,
Vermeulen
S.
,
van Roy
F.
,
Adamson
E. D.
,
Takeichi
M.
(
1998
).
Alpha-catenin-vinculin interaction functions to organize the apical junctional complex in epithelial cells.
J. Cell Biol
142
,
847
857
Weiss
E. E.
,
Kroemker
M.
,
Rudiger
A. H.
,
Jockusch
B. M.
,
Rudiger
M.
(
1998
).
Vinculin is part of the cadherin-catenin junctional complex: complex formation between alpha-catenin and vinculin.
J. Cell Biol
141
,
755
764
Wiedemann
P.
,
Giehl
K.
,
Almo
S. C.
,
Fedrov
A. A.
,
Steinberger
P.
,
Rudiger
M.
,
Ortner
M.
,
Sippl
M.
,
Dolecek
C.
,
Kraft
D.
,
Jockusch
B. M.
,
Valenta
R.
(
1996
).
Molecular and structural analysis of a continous birch profilin epitope defined by a monoclonal antibody.
J. Biol. Chem
271
,
29915
29921
Yap
A. S.
,
Brieher
W. M.
,
Pruschy
M.
,
Gumbiner
B. M.
(
1997
).
Lateral clustering of the adhesive ectodomain- a fundamental determination of cadherin function.
Curr. Biol
7
,
308
315
Yap
A. S.
,
Niessen
C. M.
,
Gumbiner
B. M.
(
1998
).
The juxtamembrane region of cadherin cytoplasmic tail supports lateral clustering, adhesive strengthening, and interaction with p120(ctn).
J. Cell Biol
4
,
779
789
Yamagishi
H.
,
Garg
V.
,
Matsuoka
R.
,
Thomas
T.
,
Srivastava
D.
(
1999
).
A molecular pathway revealing a genetic basis for human cardiac and craniofacial defects.
Science
283
,
1158
1161
This content is only available via PDF.