The IL-6 receptor complex is expressed in different polarized epithelial cells such as liver hepatocytes and intestinal cells. It consists of two subunits: gp80, which binds the ligand, and gp130, which is responsible for signal transduction. In stably transfected Madin-Darby canine kidney (MDCK) cells we have studied the localization of the human IL-6 receptor subunits and found that gp80 and gp130 are predominantly expressed at the basolateral membrane. Analysis of MDCK cells expressing truncated forms of gp80 or gp130 showed that loss of the cytoplasmic domains results in apical delivery. Expression of deletion mutants of gp80 in MDCK cells led to the identification of two discontinous motifs responsible for basolateral sorting: a membrane-proximal tyrosine-based motif (YSLG) and a more membrane-distal dileucine-type motif (LI). Activation of signal transducer and activator of transcription-3 (STAT-3) only occurred via basolaterally located gp80, suggesting that endogenous gp130 is also constrained to the basolateral plasma membrane. Our identification of a basolateral sorting signal within the cytoplasmic region of gp80 for the first time attributes a function to this domain.

REFERENCES

Andrews
N. C.
,
Faller
D. V.
(
1991
).
A rapid micropreparation technique for extraction of DNA-binding proteins from limiting numbers of mammalian cells.
Nucleic Acids Res
19
,
2499
–.
Beau
I.
,
Groyer-Picard
M. T.
,
Le Bivic
A.
,
Vannier
B.
,
Loosfelt
H.
,
Milgrom
E.
,
Misrahi
M.
(
1998
).
The basolateral localization signal of the follicle-stimulating hormone receptor.
J. Biol. Chem
273
,
18610
18616
Beau
I.
,
Misrahi
M.
,
Gross
B.
,
Vannier
B.
,
Loosfelt
H.
,
Hai
M. T.
,
Pichon
C.
,
Milgrom
E.
(
1997
).
Basolateral localization and transcytosis of gonadotropin and thyrotropin receptors expressed in Madin-Darby canine kidney cells.
J. Biol. Chem
272
,
5241
5248
Behrmann
I.
,
Janzen
C.
,
Gerhartz
C.
,
Schmitz-Van de Leur
H.
,
Hermanns
H.
,
Heesel
B.
,
Graeve
L.
,
Horn
F.
,
Tavernier
J.
,
Heinrich
P. C.
(
1997
).
A single STAT recruitment module in a chimeric cytokine receptor complex is sufficient for STAT activation.
J. Biol. Chem
272
,
5269
5274
Bresciani
R.
,
Denzer
K.
,
Pohlmann
R.
,
von Figura
K.
(
1997
).
The 46 kDa mannose-6-phosphate receptor contains a signal for basolateral sorting within the 19 juxtamembrane cytosolic residues.
Biochem. J
327
,
811
818
Brewer
C. B.
,
Roth
M. G.
(
1991
).
A single amino acid change in the cytoplasmic domain alters the polarized delivery of influenza virus hemagglutinin.
J. Cell Biol
114
,
413
421
Casanova
J. E.
,
Apodaca
G.
,
Mostov
K. E.
(
1991
).
An autonomoussignal for basolateral sorting in the cytoplasmic domain of the polymeric immunoglobulin receptor.
Cell
66
,
65
75
Castell
J. V.
,
Geiger
T.
,
Gross
V.
,
Andus
T.
,
Walter
E.
,
Hirano
T.
,
Kishimoto
T.
,
Heinrich
P. C.
(
1988
).
Plasma clearance, organ distribution and target cells of interleukin-6/hepatocyte-stimulating factor in the rat.
Eur. J. Biochem
177
,
357
361
Dittrich
E.
,
Rose-John
S.
,
Gerhartz
C.
,
Mullberg
J.
,
Stoyan
T.
,
Yasukawa
K.
,
Heinrich
P. C.
,
Graeve
L.
(
1994
).
Identification of a region within the cytoplasmic domain of the interleukin-6 (IL-6) signal transducer gp130 important for ligand-induced endocytosis of the IL-6 receptor.
J. Biol. Chem
269
,
19014
19020
Dittrich
E.
,
Haft
C. R.
,
Muys
L.
,
Heinrich
P. C.
,
Graeve
L.
(
1996
).
A di-leucine motif and an upstream serine in the interleukin-6 (IL-6) signal transducer gp130 mediate ligand-induced endocytosis and down-regulation of the IL-6 receptor.
J. Biol. Chem
271
,
5487
5494
Eaton
S.
,
Simons
K.
(
1995
).
Apical, basal, and lateral cues for epithelial polarization.
Cell
82
,
5
8
El Nemer
W.
,
Colin
Y.
,
Bauvy
C.
,
Codogno
P.
,
Fraser
R. H.
,
Cartron
J. P.
,
Le Van Kim
C. L.
) (
1999
).
Isoforms of the Lutheran/basal cell adhesion molecule glycoprotein are differentially delivered in polarized epithelial cells. Mapping of the basolateral sorting signal to a cytoplasmic di-leucine motif.
J. Biol. Chem
274
,
31903
31908
Fölsch
H.
,
Ohno
H.
,
Bonifacino
J. S.
,
Mellman
I.
(
1999
).
A novel clathrin adaptor complex mediates basolateral targeting in polarized epithelial cells [see comments].
Cell
99
,
189
198
Fuller
S. D.
,
Simons
K.
(
1986
).
Transferrin receptor polarity and recycling accuracy in ‘tight’ and ‘leaky’ strains of Madin-Darby canine kidney cells.
J. Cell Biol
103
,
1767
1779
Geffen
I.
,
Fuhrer
C.
,
Leitinger
B.
,
Weiss
M.
,
Huggel
K.
,
Griffiths
G.
,
Spiess
M.
(
1993
).
Related signals for endocytosis and basolateral sorting of the asialoglycoprotein receptor.
J. Biol. Chem
268
,
20772
20777
Gerhartz
C.
,
Dittrich
E.
,
Stoyan
T.
,
Rose-John
S.
,
Yasukawa
K.
,
Heinrich
P. C.
,
Graeve
L.
(
1994
).
Biosynthesis and half-life of the interleukin-6 receptor and its signal transducer gp130.
Eur. J. Biochem
223
,
265
274
Gerhartz
C.
,
Heesel
B.
,
Sasse
J.
,
Hemmann
U.
,
Landgraf
C.
,
Schneider-Mergener
J.
,
Horn
F.
,
Heinrich
P. C.
,
Graeve
L.
(
1996
).
Differential activation of acute phase response factor/STAT3 and STAT1 via the cytoplasmic domain of the interleukin 6 signal transducer gp130. I. Definition of a novel phosphotyrosine motif mediating STAT1 activation.
J. Biol. Chem
271
,
12991
12998
Graeve
L.
,
Drickamer
K.
,
Rodriguez-Boulan
E.
(
1989
).
Polarized endocytosis by Madin-Darby canine kidney cells transfected with functional chicken liver glycoprotein receptor.
J. Cell Biol
109
,
2809
2816
Graeve
L.
,
Korolenko
T. A.
,
Hemmann
U.
,
Weiergräber
O.
,
Dittrich
E.
,
Heinrich
P. C.
(
1996
).
A complex of the soluble interleukin-6 receptor and interleukin-6 is internalized via the signal transducer gp130.
FEBS Lett
399
,
131
134
Graeve
L.
,
Patzak
A.
,
Drickamer
K.
,
Rodriguez-Boulan
E.
(
1990
).
Polarized expression of functional rat liver asialoglycoprotein receptor in transfected Madin-Darby canine kidney cells.
J. Biol. Chem
265
,
1216
1224
Graham
F. L.
,
van der Eb
A. J.
) (
1973
).
A new technique for the assay of infectivity of human adenovirus 5 DNA.
Virology
52
,
456
467
Gut
A.
,
Kappeler
F.
,
Hyka
N.
,
Balda
M. S.
,
Hauri
H. P.
,
Matter
K.
(
1998
).
Carbohydrate-mediated Golgi to cell surface transport and apical targeting of membrane proteins.
EMBO J
17
,
1919
1929
Heinrich
P. C.
,
Behrmann
I.
,
Muller-Newen
G.
,
Schaper
F.
,
Graeve
L.
(
1998
).
Interleukin-6-type cytokine signalling through the gp130/Jak/STAT pathway.
Biochem. J
334
,
297
314
Heinrich
P. C.
,
Castell
J. V.
,
Andus
T.
(
1990
).
Interleukin-6 and the acute phase response.
Biochem. J
265
,
621
636
Hibi
M.
,
Murakami
M.
,
Saito
M.
,
Hirano
T.
,
Taga
T.
,
Kishimoto
T.
(
1990
).
Molecular cloning and expression of an IL-6 signal transducer, gp130.
Cell
63
,
1149
1157
Hunziker
W.
,
Fumey
C.
(
1994
).
A di-leucine motif mediates endocytosis and basolateral sorting of macrophage IgG Fc receptors in MDCK cells.
EMBO J
13
,
2963
2967
Koivisto
U. M.
,
Viikari
J. S.
,
Kontula
K.
(
1995
).
Molecular characterization of minor gene rearrangements in Finnish patients with heterozygous familial hypercholesterolemia: identification of two common missense mutations (Gly823Asp and Leu380 His) and eight rare mutations of the LDL receptor gene.
Am. J. Hum. Genet
57
,
789
797
Mackiewicz
A.
,
Schooltink
H.
,
Heinrich
P. C.
,
Rose-John
S.
(
1992
).
Complex of soluble human IL-6-receptor/IL-6 up-regulates expression of acute-phase proteins.
J. Immunol
149
,
2021
2027
Maratos-Flier
E.
,
Kao
C. Y.
,
Verdin
E. M.
,
King
G. L.
(
1987
).
Receptor-mediated vectorial transcytosis of epidermal growth factor by Madin-Darby canine kidney cells.
J. Cell Biol
105
,
1595
1601
Matter
K.
,
Hunziker
W.
,
Mellman
I.
(
1992
).
Basolateral sorting of LDL receptor in MDCK cells: the cytoplasmic domain contains two tyrosine-dependent targeting determinants.
Cell
71
,
741
753
Matter
K.
,
Mellman
I.
(
1994
).
Mechanisms of cell polarity: sorting and transport in epithelial cells.
Curr. Opin. Cell Biol
6
,
545
554
Matter
K.
,
Whitney
J. A.
,
Yamamoto
E. M.
,
Mellman
I.
(
1993
).
Common signals control low density lipoprotein receptor sorting in endosomes and the Golgi complex of MDCK cells.
Cell
74
,
1053
1064
Molmenti
E. P.
,
Ziambaras
T.
,
Perlmutter
D. H.
(
1993
).
Evidence for an acute phase response in human intestinal epithelial cells.
J. Biol. Chem
268
,
14116
14124
Monlauzeur
L.
,
Rajasekaran
A.
,
Chao
M.
,
Rodriguez-Boulan
E.
,
Le Bivic
A.
(
1995
).
A cytoplasmic tyrosine is essential for the basolateral localization of mutants of the human nerve growth factor receptor in Madin-Darby canine kidney cells.
J. Biol. Chem
270
,
12219
12225
Muller-Newen
G.
,
Köhne
C.
,
Keul
R.
,
Hemmann
U.
,
Muller-Esterl
W.
,
Wijdenes
J.
,
Brakenhoff
J. P.
,
Hart
M. H.
,
Heinrich
P. C.
(
1996
).
Purification and characterization of the soluble interleukin-6 receptor from human plasma and identification of an isoform generated through alternative splicing.
Eur. J. Biochem
236
,
837
842
Ogborn
M. R.
,
Sareen
S.
,
Tomobe
K.
,
Takahashi
H.
,
Crocker
J. F.
(
1995
).
Renal tubule Na,K-ATPase polarity in different animal models of polycystic kidney disease.
J. Histochem. Cytochem
43
,
785
790
Ohno
H.
,
Tomemori
T.
,
Nakatsu
F.
,
Okazaki
Y.
,
Aguilar
R. C.
,
Fölsch
H.
,
Mellman
I.
,
Saito
T.
,
Shirasawa
T.
,
Bonifacino
J. S.
(
1999
).
1B, a novel adaptor medium chain expressed in polarized epithelial cells.
FEBS Lett
449
,
215
220
Prydz
K.
,
Brandli
A. W.
,
Bomsel
M.
,
Simons
K.
(
1990
).
Surface distribution of the mannose 6-phosphate receptors in epithelial Madin-Darby canine kidney cells.
J. Biol. Chem
265
,
12629
12635
Rapoport
I.
,
Chen
Y. C.
,
Cupers
P.
,
Shoelson
S. E.
,
Kirchhausen
T.
(
1998
).
Dileucine-based sorting signals bind to the beta chain of AP-1 at a site distinct and regulated differently from the tyrosine-based motif-binding site.
EMBO J
17
,
2148
2155
Rodionov
D. G.
,
Bakke
O.
(
1998
).
Medium chains of adaptor complexes AP-1 and AP-2 recognize leucine-based sorting signals from the invariant chain.
J. Biol. Chem
273
,
6005
6008
Rodriguez-Boulan
E.
,
Powell
S. K.
(
1992
).
Polarity of epithelial and neuronal cells.
Annu. Rev. Cell Biol
8
,
395
427
Rodriguez-Boulan
E.
,
Salas
P. J.
,
Sargiacomo
M.
,
Lisanti
M.
,
Le Bivic
A.
,
Sambuy
Y.
,
Vega-Salas
D.
,
Graeve
L.
(
1989
).
Methods to estimate the polarized distribution of surface antigens in cultured epithelial cells.
Methods Cell Biol
32
,
37
56
Sanger
F.
,
Nicklen
S.
,
Coulson
A. R.
(
1977
).
DNA sequencing with chain-terminating inhibitors.
Proc. Natl. Acad. Sci. USA
74
,
5463
5467
Simons
K.
,
Fuller
S. D.
(
1985
).
Cell surface polarity in epithelia.
Annu. Rev. Cell Biol
1
,
243
288
Simonsen
A.
,
Bremnes
B.
,
Nordeng
T. W.
,
Bakke
O.
(
1998
).
The leucine-based motif DDQxxLI is recognized both for internalization and basolateral sorting of invariant chain in MDCK cells.
Eur. J. Cell. Biol
76
,
25
32
Sonne
O.
,
Davidsen
O.
,
Moller
B. K.
,
Munck Petersen
C.
(
1990
).
Cellular targets and receptors for interleukin-6. I. In vivo and in vitro uptake of IL-6 in liver and hepatocytes. Eur.
J. Clin. Invest
20
,
366
376
Stahl
N.
,
Farruggella
T. J.
,
Boulton
T. G.
,
Zhong
Z.
,
Darnell
J. E.
Jr.
,
Yancopoulos
G. D.
(
1995
).
Choice of STATs and other substrates specified by modular tyrosine-based motifs in cytokine receptors.
Science
267
,
1349
1353
Sweeney
W. E.
Jr.
,
Avner
E. D.
(
1998
).
Functional activity of epidermal growth factor receptors in autosomal recessive polycystic kidney disease.
Am. J. Physiol
275
,
387
394
Taga
T.
,
Hibi
M.
,
Hirata
Y.
,
Yamasaki
K.
,
Yasukawa
K.
,
Matsuda
T.
,
Hirano
T.
,
Kishimoto
T.
(
1989
).
Interleukin-6 triggers theassociation of its receptor with a possible signal transducer, gp130.
Cell
58
,
573
581
van Meer
G.
,
Stelzer
E. H.
,
Wijnaendts-van-Resandt
R. W.
,
Simons
K.
(
1987
).
Sorting of sphingolipids in epithelial (Madin-Darby canine kidney) cells.
J. Cell Biol
105
,
1623
1635
Wagner
B. J.
,
Hayes
T. E.
,
Hoban
C. J.
,
Cochran
B. H.
(
1990
).
The SIF binding element confers sis/PDGF inducibility onto the c-fos promoter.
EMBO J
9
,
4477
4484
Wegenka
U. M.
,
Buschmann
J.
,
Lutticken
C.
,
Heinrich
P. C.
,
Horn
F.
(
1993
).
Acute-phase response factor, a nuclear factor binding to acute-phase response elements, is rapidly activated by interleukin-6 at the posttranslational level.
Mol. Cell. Biol
13
,
276
288
Wessels
H. P.
,
Geffen
I.
,
Spiess
M.
(
1989
).
A hepatocyte-specific basolateral membrane protein is targeted to the same domain when expressed in Madin-Darby canine kidney cells.
J. Biol. Chem
264
,
17
20
Wijdenes
J.
,
Heinrich
P. C.
,
Muller-Newen
G.
,
Roche
C.
,
Gu
Z. J.
,
Clement
C.
,
Klein
B.
(
1995
).
Interleukin-6 signal transducer gp130 has specific binding sites for different cytokines as determined byantagonistic and agonistic anti-gp130 monoclonal antibodies.
Eur. J. Immunol
25
,
3474
3481
Wozniak
M.
,
Limbird
L. E.
(
1996
).
The three alpha 2-adrenergic receptor subtypes achieve basolateral localization in Madin-Darby canine kidney II cells via different targeting mechanisms.
J. Biol. Chem
271
,
5017
5024
Yamasaki
K.
,
Taga
T.
,
Hirata
Y.
,
Yawata
H.
,
Kawanishi
Y.
,
Seed
B.
,
Taniguchi
T.
,
Hirano
T.
,
Kishimoto
T.
(
1988
).
Cloning and expression of the human interleukin-6 (BSF-2/IFN beta 2) receptor.
Science
241
,
825
828
Yokode
M.
,
Pathak
R. K.
,
Hammer
R. E.
,
Brown
M. S.
,
Goldstein
J. L.
,
Anderson
R. G.
(
1992
).
Cytoplasmic sequence required for basolateral targeting of LDL receptor in livers of transgenic mice.
J. Cell. Biol
117
,
39
46
Zohlnhöfer
D.
,
Graeve
L.
,
Rose-John
S.
,
Schooltink
H.
,
Dittrich
E.
,
Heinrich
P. C.
(
1992
).
The hepatic interleukin-6 receptor. Down-regulation of the interleukin-6 binding subunit (gp80) by its ligand.
FEBS Lett
306
,
219
222
This content is only available via PDF.