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J Biol Chem, Vol. 273, Issue 25, 15719-15726, June 19, 1998
From the Growth hormone (GH) and
prolactin (PRL) binding to their receptors, which belong to the
cytokine receptor superfamily, activate Janus kinase (JAK) 2 tyrosine
kinase, thereby leading to their biological actions. We recently showed
that GH mainly stimulated tyrosine phosphorylation of epidermal growth
factor receptor and its association with Grb2, and concomitantly
stimulated mitogen-activated protein kinase activity in liver, a major
target tissue. Using specific antibodies, we now show that GH was also
able to induce tyrosine phosphorylation of insulin receptor substrate
(IRS)-1/IRS-2 in liver. In addition, the major tyrosine-phosphorylated
protein in anti-p85 phosphatidylinositol 3-kinase (PI3-kinase)
immunoprecipitate from liver of wild-type mice was IRS-1, and IRS-2 in
IRS-1 deficient mice, but not epidermal growth factor receptor. These
data suggest that tyrosine phosphorylation of IRS-1 may be a major
mechanism for GH-induced PI3-kinase activation in physiological target
organ of GH, liver. We also show that PRL was able to induce tyrosine phosphorylation of both IRS-1 and IRS-2 in COS cells transiently transfected with PRLR and in CHO-PRLR cells. Moreover, we show that
tyrosine phosphorylation of IRS-3 was induced by both GH and PRL in COS
cells transiently transfected with IRS-3 and their cognate receptors.
By using the JAK2-deficient cell lines or by expressing a
dominant negative JAK2 mutant, we show that JAK2 is required for the
GH- and PRL-dependent tyrosine phosphorylation of IRS-1,
-2, and -3. Finally, a specific PI3-kinase inhibitor, wortmannin,
completely blocked the anti-lipolytic effect of GH in 3T3 L1
adipocytes. Taken together, the role of IRS-1, -2, and -3 in GH and PRL
signalings appears to be phosphorylated by JAK2, thereby providing
docking sites for p85 PI3-kinase and activating PI3-kinase and its
downstream biological effects.
Growth hormone (GH)1 and
prolactin (PRL) initiate their wide variety of biological effects by
binding and dimerization of their membrane receptors (1, 2). The GH and
PRL receptors belong to the cytokine/hematopoietin receptor
superfamily, characterized by homologies in the extracellular domains
and lack of intrinsic tyrosine kinase activity (1), however, ligands
binding to their receptors activate JAK2 tyrosine kinase (3, 4). JAK2
tyrosine phosphorylates their receptors and JAK2 itself along with
STATs (signal transducers and activators of transcription). The
phosphorylated STAT proteins translocate into the nucleus and bind to
DNA, thereby activating transcription of specific genes (5). In
addition, a number of intracellular key proteins have been suggested to be involved in their signaling (6).
One of the signaling molecules known to be activated by GH and PRL (7,
8) is the mitogen-activated protein kinase (MAP kinase), which is
believed to play a pivotal role in the regulation of cellular growth
and differentiation (9). Association of tyrosine-phosphorylated
proteins with Grb2 (growth factor receptor bound protein 2) is known to
represent a crucial step in the activation of MAP kinase cascade (10).
In the case of GH and PRL, epidermal growth factor receptor (EGFR) and
Shc have been shown to be tyrosyl phosphorylated by JAK2 and bind Grb2,
leading to MAP kinase activation (11, 12).
Another signaling molecule known to be activated by GH and PRL is
phosphatidylinositol 3-kinase (PI3-kinase), which may play a role in
initiating insulin-like effects of GH and PRL including glucose
transport, glycogenesis, anti-lipolysis, and lipogenesis (13-15),
because the selective PI3-kinase inhibitor wortmannin can block at
least some of these effects such as anti-lipolysis and lipogenesis
(16).
Insulin receptor substrate-1 (IRS-1) is the principal substrate of the
insulin receptor kinase whose molecular mass is approximately 170 kDa
and has many tyrosine phosphorylation sites (17), which provides
binding sites for several distinct Src homology 2 (SH2) proteins
(e.g. Grb2, the 85-kDa subunit of PI3-kinase (PI3-kinase p85), Syp, Nck, and Csk) and has been shown to mediate multiple signaling pathways (18, 19). IRS-1 binds the PI3-kinase p85 when
tyrosine phosphorylated, thereby activating PI3-kinase (20, 21),
leading to exert metabolic effects of insulin such as glucose transport, glycogen synthesis, and anti-lipolysis (22-24).
To better understand the role of IRS-1 in vivo, we and
others generated mice with a targeted disruption of the IRS-1 gene and
demonstrated that they exhibited mild growth retardation and had
partial resistance to the glucose-lowering effect of insulin, which was
an unexpectedly mild phenotype, suggesting the presence of IRS-1
independent pathways (25, 26). In liver and also muscle extracts from
IRS-1-deficient mice, tyrosine phosphorylation of IRS-2, whose
molecular mass is approximately 180 kDa (25, 27, 28), was significantly
induced in IRS-1-deficient mice compared with that in wild-type mice
(29) and has been suggested to be the mechanism of compensation for
IRS-1 deficiency in liver and muscle (30, 31). In another target tissue
of insulin, adipocytes, both IRS-1 and IRS-3 (pp60) (32, 33) was
suggested to play a major role in insulin-induced PI3-kinase
activation, and IRS-3 (pp60) was suggested to be involved in regulating
this process in the absence of IRS-1 (34).
Recently, IRS-1/IRS-2 have been shown to be tyrosine-phosphorylated and
associated with PI3-kinase p85 in response to GH in primary rat
adipocytes (35, 36) and in 3T3-F442A fibroblasts (37, 38). However, the
following important issues remained unresolved. First, whether GH was
able to induce tyrosine phosphorylation of IRS-1/IRS-2 in physiological
target organ, liver, in vivo. Second, the relative
contributions of IRS-1 and IRS-2 in the signaling pathways for GH to
activate PI3-kinase. Third, the identity of the kinase responsible for
the GH-stimulated tyrosine phosphorylation of IRS proteins. Fourth,
whether additional cytokine receptor superfamily/IRS proteins signaling
pathways could also operate. Using specific antibodies, we show here
that GH was able to induce tyrosine phosphorylation of IRS-1/IRS-2, and
that tyrosine phosphorylation of IRS-1 may be a major mechanism for
GH-induced PI3-kinase activation in physiological target organ, liver,
using IRS-1-deficient mice. Finally, by using the JAK2-deficient cell
lines or by expressing a dominant negative JAK2 mutant, we show that
JAK2 is required for the GH- and PRL-dependent tyrosine
phosphorylation of IRS-1, -2, and -3, their association with p85
PI3-kinase and activation of PI3-kinase. Taken together, the role of
IRS-1, -2, and -3 in GH and PRL signalings is to be phosphorylated by
JAK2, thereby providing docking sites for p85 PI3-kinase and activating
PI3-kinase. A specific PI3-kinase inhibitor, wortmannin, completely
blocked the anti-lipolytic effect of GH in 3T3 L1 adipocytes, as
reported in isolated rat adipocytes (16). Thus our data may provide the biochemical fundamentals to understand the insulin-like effects of GH
such as anti-lipolysis.
Materials and Mice--
The polyclonal antibody against 85-kDa
subunit of PI3-kinase was from Upstate Biotechnology Inc. The
monoclonal antibody against hemagglutinin (HA), 12CA5, was from
Boehringer Mannheim. All other materials were obtained from the sources
described (12, 31). Mice were fasted overnight and then treated as
described (31).
Expression of Wild-type and Kinase-inactive JAK2, Epitope-tagged
IRS-3, GHR, and PRLR--
The CHO (Chinese hamster ovary)-PRLR cells
(approximately 1.0 × 104 receptors/cell) and
expression vector of human PRLR were kindly provided by M. Takahashi,
M. Wada, and M. Honjo. Immunoprecipitation, Immunoblotting, and PI3-kinase
Activities--
Immunoprecipitation and immunoblotting were performed
as described (31). The amount of tyrosine phosphorylation or associated protein was evaluated by densitometry, as described (31). The significance of differences between the two groups was assessed using
Student's unpaired t test.
Lipolysis--
3T3-L1 adipocytes were incubated in serum-free
Dulbecco's modified Eagle's medium for 16 h, and then in
modified Krebs-Ringer medium at pH 7.5 containing 24 mM
HEPES, 119 mM NaCl, 4.95 mM KCl, 2.54 mM CaCl2, 1.19 mM
KH2PO4, 1.19 mM MgSO4,
2 mM glucose for 3 h. Lipolysis, stimulated by
norepinephrine (NE) (Sigma), was measured with or without pretreatment
with 100 nM wortmannin for 20 min, as proportional to
glycerol released to the medium during a 30-min incubation and
determined as described (16).
GH Stimulates Tyrosine Phosphorylation of IRS-1/IRS-2 in Liver, in
Vivo--
To determine if IRS-1 and/or IRS-2 are
tyrosine-phosphorylated following stimulation with GH in its
physiological target organ, liver, in vivo, mice were
injected with 5 µg/g body weight of GH and 7.5 min after the
injection, solubilized proteins from livers of wild-type and IRS-1
deficient mice were immunoprecipitated with anti-IRS-1 antibody
(
Growth Hormone and Prolactin Stimulate Tyrosine Phosphorylation
of Insulin Receptor Substrate-1, -2, and -3, Their Association with p85
Phosphatidylinositol 3-Kinase (PI3-kinase), and Concomitantly
PI3-kinase Activation via JAK2 Kinase*
,,,,
,,,, and
Third Department of Internal Medicine,
Faculty of Medicine, University of Tokyo, Tokyo 113, Japan, the
§ Department of Molecular Biology, Research Institute, The
Cleveland Clinic Foundation, Cleveland, Ohio 44195, the
¶ Imperial Cancer Research Fund, London WC2A 3PX, United Kingdom,
the Second Department of Internal Medicine, Tokyo Women's
Medical College, Tokyo 162, Japan, and the ** Institute for Diabetes
Care and Research, Asahi Life Foundation, Tokyo 100, Japan
ABSTRACT
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Abstract
Introduction
Procedures
Results
Discussion
References
INTRODUCTION
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Abstract
Introduction
Procedures
Results
Discussion
References
EXPERIMENTAL PROCEDURES
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Introduction
Procedures
Results
Discussion
References
2A/GHR (39), CHO-GHR, or CHO-PRLR cells were
transfected with or without 3 µg of WT-JAK2 plasmids or dominant
negative JAK2 (
JAK2) (40) that lacks the C terminus kinase domain
under the SR
promoter in 6-cm dishes by the LipofectAMINE method
with modification and the cells were stimulated as described (12, 39,
41). The rat IRS-3 cDNA in the pcDNA3 was modified to encode an
additional 9-amino acid HA epitope (YPYDVPDYA), after the C terminus of
IRS-3 via a recombinant polymerase chain reaction strategy (33). COS cells were transfected with 3 µg of GHR or PRLR, with or without HA-tagged IRS-3 and with or without 2 µg of WT-JAK2 or JAK2 at 60-80% confluence in 3 ml of Dulbecco's modified Eagle's medium in
6-cm dishes by the calcium phosphate precipitation method and the cells
were stimulated as described (12).
RESULTS
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Abstract
Introduction
Procedures
Results
Discussion
References
IRS-1) or anti-IRS-2 antibody (IRS-2), and immunoblotted with
PY. IRS-1 (molecular mass 170 kDa) was tyrosine phosphorylated in
response to GH in wild-type mice (Fig. 1A, lane 3),
however, to a much lesser degree than in response to insulin (Fig.
1A, lane 2). IRS-2 was tyrosine phosphorylated in response
to GH most in IRS-1 deficient mice (Fig. 1B, lane 6),
however, to a much lesser degree than in response to insulin (Fig.
1B, lane 5). GH-dependent tyrosine
phosphorylation of IRS-1/IRS-2 reached the maximum level at 7.5 min
(data not shown).
View larger version (42K):
[in a new window]
Fig. 1.
GH-stimulated tyrosine phosphorylation of
IRS-1/IRS-2 in livers from wild-type (WT) and IRS-1
deficient mice (null). A and B,
upper panel, liver extracts from wild-type or IRS-1
deficient mice untreated or treated with insulin for 2 min or treated
with GH for 7.5 min were subjected to immunoprecipitation with IRS-1
(A) or IRS-2 (B) followed by Western blotting
with PY. A and B, lower panel,
amount of tyrosine phosphorylation of IRS-1 (A) or IRS-2
(B) in livers. The amount of tyrosine phosphorylation was
evaluated by densitometry and expressed as the percentage of the value
of wild-type mice treated with insulin. Each bar represents
the mean ± S.E. for livers from 5 to 7 mice. *, p < 0.05, N.S., the difference is not significant, GH
injected mice versus untreated mice.
GH-stimulated PI3-kinase Activity in Liver-- We measured PI3-kinase activity in livers from wild-type and IRS-1-deficient mice in the immunoprecipitates with the monoclonal antibody against phosphotyrosine (4G10). 7.5 min after GH injection and 2 min after insulin injection, maximal levels in kinase activity were reached (data not shown). In liver of both wild-type and IRS-1-deficient mice, insulin injection resulted in a 12-13-fold increase in PI-3 kinase activity in 4G10 immunoprecipitates, while GH caused an only 2-3-fold increase (Fig. 2).
|
) or IRS-1-deficient mice (
) untreated or
treated with insulin for 2 min or treated with GH for 7.5 min were
subjected to PI3-kinase assay. The autoradiogram of the thin layer
chromatograph is shown in the left panel. The radioactivity
in the spots corresponding to phosphatidylinositol (PIP) was
measured and the results are shown in the right panel,
expressed as the ratio to the value of untreated wild-type mice. Each
bar represents the mean ± S.E. for livers from five
mice. *, p < 0.05, GH injected mice versus
untreated mice.
Immunological Detection of GH-stimulated Tyrosine-phosphorylated
Proteins Associated with PI3-kinase p85--
The molecular mechanism
by which GH activates PI3-kinase were determined by immunoblotting with
the monoclonal anti-phosphotyrosine antibody (4G10) of anti-PI3-kinase
p85 immunoprecipitates of livers 7.5 min after GH injection and 2 min
after insulin injection. Interestingly, while the 180-kDa protein was
mainly tyrosine phosphorylated and associated with Grb2 in wild-type
mice when injected with GH (12), in p85 immunoprecipitates of
GH-injected wild-type mice, a 170-kDa tyrosine-phosphorylated protein
was mainly present (Fig. 3A, lane
4), indicating that tyrosine phosphorylation of IRS-1, but neither
IRS-2 nor EGFR, may be a major mechanism for GH-induced PI3-kinase
activation in physiological target organ, liver. In wild-type mice, the
amount of pp170 associated with PI3-kinase p85 in response to GH was
much less than that in response to insulin, which was consistent with
that the GH-stimulated tyrosine phosphorylation of IRS-1 was much less
than that stimulated with insulin (Fig. 1A, lanes 2 and 3). To confirm these observations, liver extracts
of GH-treated wild-type mice for 7.5 min were incubated with the
glutathione S-transferase fusion proteins containing the
entire p85 PI3-kinase. Glutathione S-transferase-p85 only associated with pp170 (IRS-1) (data not shown). In IRS-1-deficient mice, the amount of pp180 (IRS-2) associated with PI3-kinase p85 in
response to GH was also much less than that in response to insulin
(Fig. 3A, lanes 5 and 6).
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IRS-1) or anti-IRS-2 antibody (IRS-2 (179C)). In liver of
wild-type mice, insulin injection resulted in a 11-fold increase of
PI-3 kinase activity in IRS-1 immunoprecipitates, while GH caused
only a 2-3-fold increase (Fig. 3B, lanes 2 and
3), which was consistent with the data of the amount of PI3-kinase
p85-associated pp170 (IRS-1) (Fig. 3A, lanes 3 and
4). In IRS-2 immunoprecipitates from liver of wild-type mice,
however, GH had little effect on an increase in PI3-kinase, and in
IRS-1 deficient mice, GH caused only a 2-3-fold increase (Fig.
3C, lanes 2 and 4), which was also consistent
with the data of the amount of pp180 (IRS-2) associated with PI3-kinase
p85 (Fig. 3A, lanes 4 and 6).
Complementation with a WT-JAK2 Expression Plasmid into
JAK2-deficient Cell Line Restored GH-dependent Tyrosine
Phosphorylation of IRS-1/IRS-2, Their Association with p85 PI3-kinase,
and Activation of PI3-kinase--
We next examined whether GH-induced
tyrosine phosphorylation of IRS-1/IRS-2, IRS-1 association with p85
PI3-kinase, and PI3-kinase activation are dependent on JAK2 by using
2A/GHR cells, which lack JAK2 (39). GH was unable to induce these
responses in 2A/GHR, whereas complementation with a WT-JAK2
expression plasmid restored them (Fig.
4). These data suggested that JAK2 is
required for the GH-dependent tyrosine phosphorylation of
IRS-1 and -2, their association with p85 PI3-kinase, and activation of
PI3-kinase.
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1). Upon lysis,
precipitated IRS-1 (A, upper panel) or IRS-2 (C, upper
panel) was immunoblotted with PRL Was Also Able to Induce Tyrosine Phosphorylation of IRS-2-- Recently, IRS-1 also has been shown to be tyrosine phosphorylated in response to another cytokine superfamily member PRL in 293-PRL receptor cells (42). Thus we next examined whether PRL was able to induce tyrosine phosphorylation of IRS-2 by immunoblotting with the monoclonal anti-phosphotyrosine antibody (4G10) of anti-IRS-2 immunoprecipitates of COS cells transiently transfected with PRLR and CHO-PRLR cells treated with PRL for 5 min. We found that PRL was also able to induce tyrosine phosphorylation of IRS-2 (Fig. 5D, lane 2, and Fig. 6C, lane 6).
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Complementation with a WT-JAK2 Expression Plasmid into
JAK2-deficient Cell Line Restored PRL-dependent Tyrosine
Phosphorylation of IRS-1/IRS-2, Their Association with p85 PI3-kinase,
and Activation of PI3-kinase--
We also examined whether PRL-induced
tyrosine phosphorylation of IRS-1/IRS-2, IRS-1 association with p85
PI3-kinase, and PI3-kinase activation are dependent on JAK2 by using
COS cells, in which the level of expression of JAK2 appears to be
sufficiently low that some level of coexpression of wild-type (WT)-JAK2
and PRLR is required in order to detect activation of JAK2 by PRL (Fig. 5A). Neither expression of PRLR alone nor coexpression of
kinase-inactive JAK2 (JAK2) and PRLR, but coexpression of WT-JAK2
and PRLR restored these responses (Fig. 5B, C, D, and
E, and data not shown). These data suggested that JAK2 is required
for the PRL-dependent tyrosine phosphorylation of IRS-1 and
-2, their association with p85 PI3-kinase and activation of
PI3-kinase.
Dominant Negative JAK2 Almost Completely Abolished GH- and
PRL-dependent Tyrosine Phosphorylation of IRS-1/IRS-2,
Their Association with p85 PI3-kinase, and Activation of
PI3-kinase--
We also used a dominant negative JAK2 mutant, since
the possibility of overestimation of the complemented molecule (JAK2) cannot be excluded in the above experiment. Expression of JAK2, which inhibits autophosphorylation of WT-JAK2 in a dominant negative manner (40), almost completely abolished the GH- and PRL-induced tyrosine phosphorylation of IRS-1/IRS-2, IRS-1 association with p85
PI3-kinase, and activation of PI3-kinase in CHO cells expressing their
cognate receptors (Fig. 6 and data not shown). These data suggested
that the GH- and PRL-induced tyrosine phosphorylation of IRS-1/IRS-2,
IRS-1 association with p85 PI3-kinase, and activation of PI3-kinase are
largely dependent on JAK2.
GH and PRL Were Also Able to Induce Tyrosine Phosphorylation of IRS-3 and Its Association with p85 PI3-kinase via JAK2-- Not only IRS-1 but also pp60 was shown to play a major role in insulin-induced activation of PI3-kinase in adipocytes (34). Recently, Lavan et al. (33) reported the isolation of the cDNA of pp60 which was designated as IRS-3. Thus we next examined whether GH and PRL were able to induce tyrosine phosphorylation of IRS-3 and its association with p85 PI3-kinase via JAK2 in COS cells transiently transfected with HA-tagged IRS-3 treated with GH or PRL for 5 min. We found that both GH and PRL were also able to induce tyrosine phosphorylation of IRS-3 and its association with p85 PI3-kinase via JAK2 (Fig. 7, A and B).
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GH Effect on Anti-lipolysis Is Dependent on PI3-kinase-- To study whether GH stimulation of PI3-kinase is linked to GH-induced insulin-like metabolic effect, GH-stimulated anti-lipolysis was measured with pretreatment with a specific PI3-kinase inhibitor, wortmannin. The anti-lipolytic effect of GH was measured as the ability to counteract lipolysis induced by NE at 100 nM in 3T3 L1 adipocytes. GH (0.5 µg/ml) inhibited lipolysis by 60 ± 10% (mean ± S.E., n = 3, p < 0.05), and wortmannin completely blocked this anti-lipolytic effect, as reported in isolated rat adipocytes (16). These data suggested that PI3-kinase activity was indeed required for anti-lipolysis in response to GH.
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DISCUSSION |
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Top
Abstract
Introduction
Procedures
Results
Discussion
References
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GH Stimulates Tyrosine Phosphorylation of IRS-1/IRS-2 in Liver, in Vivo-- We recently showed that GH mainly stimulated tyrosine phosphorylation of EGFR and its association with Grb2, and concomitantly stimulated MAP kinase activity in liver, a major target tissue (12). Using specific antibodies, we provided the first evidence that GH was also able to induce tyrosine phosphorylation of IRS-1/IRS-2 in physiological target organ, liver, in vivo, although to a lesser extent as compared with that stimulated with insulin or tyrosine phosphorylation of EGFR stimulated with GH (Fig. 1 and Ref. 12). The degree of GH-stimulated activation of MAP kinase and PI3-kinase appeared to be correlated with the relative amount of GH-stimulated tyrosine-phosphorylated EGFR and IRS-1, respectively (Figs. 1 and 2 and Ref. 12).
Tyrosine Phosphorylation of IRS-1, but Neither IRS-2 nor EGFR, May
Be a Major Mechanism for GH-induced PI3-kinase Activation in
Physiological Target Organ, Liver--
IRS-2 couple more
sensitively to the IL-4 receptor system and less to the insulin
receptor system than IRS-1 (28). In the case of the insulin receptor
system, the degree of compensation for IRS-1 deficiency appears to be
correlated with the relative amount of tyrosine-phosphorylated IRS-2
(in IRS-1 deficient mice) to that of IRS-1 (in wild-type mice) (31).
Recently, using specific antibodies, IRS-1/IRS-2 have been shown to be
associated with PI3-kinase p85 in response to GH in primary rat
adipocytes (35, 36) and in 3T3-F442A fibroblasts (37, 38). There has
been no study, however, which reported the relative contributions of IRS-1 and IRS-2 in the signaling pathways for GH to activate
PI3-kinase. Interestingly, although EGFR was mainly tyrosine
phosphorylated and associated with Grb2 when injected with GH (12), the
major tyrosine-phosphorylated protein in anti-p85 PI3-kinase
immunoprecipitate from liver of wild-type mice was IRS-1, and IRS-2 in
IRS-1-deficient mice (Fig. 3 A) but not EGFR. Thus we
provided the evidence that tyrosine phosphorylation of IRS-1, but
neither IRS-2 nor EGFR, may be a major mechanism for GH-induced
PI3-kinase activation in physiological target organ, liver, in
vivo. The observations that GH had little effect on an increase in
PI3-kinase activity in IRS-2 immunoprecipitates from wild-type mice,
while GH caused a 2-3-fold increase in IRS-1-deficient mice (Fig.
3C) may support the central role of IRS-1 in the control of
PI3-kinase activation by GH.
Additional Cytokine Receptor Superfamily/IRS Proteins Signaling Pathways-- Recently, PRL has been shown to be able to tyrosine phosphorylate IRS-1 in 293-PRLR cells (42). We show in this study for the first time that PRL was also able to induce tyrosine phosphorylation of IRS-2 in COS cells transiently transfected with PRLR (Fig. 5D, lane 2) and in CHO-PRLR cells (Fig. 6C, lane 6). In addition to IRS-1, pp60/IRS-3 also has been shown to play a major role in insulin-induced activation of PI3-kinase in adipocytes (34). In the present study, we also show that both GH and PRL were able to induce tyrosine phosphorylation of IRS-3 and its association with p85 PI3-kinase (Fig. 7). This is the first evidence that IRS-3 can be tyrosine phosphorylated by other tyrosine kinase(s) other than insulin receptor tyrosine kinase.
The GH- and PRL-induced Tyrosine Phosphorylation of IRS-1, -2, and -3, Their Association with p85 PI3-kinase, and Activation of PI3-kinase Are Largely Dependent on JAK2-- There has been no study reporting the identity of the kinase responsible for GH-induced tyrosine phosphorylation of IRS-1/IRS-2, although Argetsinger et al. (37, 38) reported that the regions of GHR necessary for IRS-1/IRS-2 tyrosyl phosphorylation were present between amino acids 295 and 380 containing box 1 and box 2, the same as those required for JAK2 association and tyrosyl phosphorylation. By using the JAK2-deficient cell lines or expressing a dominant negative JAK2 mutant, we provided the first direct evidence that JAK2 is required for the GH- and PRL-dependent tyrosine phosphorylation of IRS-1, -2, and -3, their association with p85 PI3-kinase, and activation of PI3-kinase (Figs. 4-7). Tyrosine phosphorylation of IRS-1 induced by insulin, insulin-like growth factor-I, and interleukin-4 appears to require interaction between the phosphotyrosine-binding domain of IRS-1 and the tyrosine-phosphorylated NPXY motif in the ligand receptor (43-45). GHR and JAK2, however, do not contain NPXY motifs (46, 47). This may explain, at least in part, why GH was able to induce tyrosine phosphorylation of IRS-1/IRS-2, their association with p85 PI3-kinase, and activation of PI3-kinase, however, to a lesser extent as compared with that stimulated with insulin. The domains required for interactions between cytokine superfamily-JAK complexes and IRS proteins are now under investigation.
JAK/IRS Family and JAK/EGFR Pathways for Cytokine Receptor
Superfamily--
An essential role of JAK/STAT pathways is now
established for cytokine receptor superfamily (5). The roles of JAKs in
the activation of additional pathways, however, have been less well established. Burfoot et al. (5) reported that tyrosine
phosphorylation of IRS-1 mediated by the Type I interferons is
dependent on JAK1 and TYK2, and by interleukin 4 and oncostatin M is
largely JAK1-dependent, by complementation with JAK into
mutant cell lines lacking JAK (41). The possibility of overestimation
of the complemented molecule, however, cannot be excluded. Thus we also
used a dominant negative JAK2 mutant. Expression of dominant negative
JAK2 almost completely abolished GH- and PRL-induced tyrosine
phosphorylation of IRS-1 and -2, their association with p85 PI3-kinase
and activation of PI3-kinase (Fig. 6), indicating that these responses
are largely dependent on JAK2. Together with others (12, 41)
observations, our findings using a dominant negative molecule provide
the paradigm of JAK/IRS family and JAK/EGFR pathways for cytokine
receptor superfamily. STAT specificity might be determined not by each JAK but in concert with the cytoplasmic structure of the particular receptor with which it is associated. In the case of IRS family, for
example, tyrosine phosphorylation of IRS-1, but not IRS-2, is dependent
on JAK1 and Tyk2 for the 
-interferons, while neither IRS-1 nor
IRS-2 was detectably tyrosine phosphorylated by interferon-, which
was able to activate JAK1 and JAK2 (41). Furthermore, we showed that
GH- and PRL-induced tyrosine phosphorylation of IRS-1, -2, and -3 are
largely dependent on JAK2 (Figs. 4-7). Thus like STAT specificity, IRS
specificity might be determined not by each JAK but by each
receptor.
The Role of PI3-kinase in Mediating Insulin-like Effects of GH-- PI3-kinase has been reported not to be required for glucose uptake stimulated by GH (48). GH-induced anti-lipolysis and lipogenesis, however, have been shown to be PI3-kinase-dependent in isolated rat adipocytes (16). We also showed that a specific PI3-kinase inhibitor, wortmannin, completely blocked the anti-lipolytic effect of GH in 3T3 L1 adipocytes (Fig. 8). Thus PI3-kinase may play an important role in initiating at least some insulin-like effects of GH, such as anti-lipolysis and lipogenesis.
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ACKNOWLEDGEMENTS |
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We thank J. Ihle for JAK2 cDNA, K. Arai
for expression vectors of JAK2 and dominant negative JAK2 (JAK2), M. Takahashi, M. Wada, and M. Honjo for CHO-PRLR and CHO-GHR cells, and S. Kakinuma for technical assistance.
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FOOTNOTES |
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* This work was supported by a grant from the Research Fellowships of the Japan Society for the Promotion of Science for Young Scientists (to T. Y.), from the Ministry of Education, Science, Sports, and Culture and the Ministry of Health and Welfare of Japan (to T. K.) and a grant for diabetes research from the Taisho Pharmaceutical Co., Ltd. (to T. K.).The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.
To whom correspondence should be addressed: Third Department of
Internal Medicine, Faculty of Medicine, University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-0033, Japan. Tel.: 81-3-3815-5411 (ext:
3111); Fax: 81-3-5689-7209; E-mail:
kadowaki-3im{at}h.u-tokyo.ac.jp.
1
The abbreviations used are: GH, growth hormone;
PRL, prolactin; JAK, Janus kinase; STAT, signal transducers and
activators of transcription; MAP, mitogen-activated protein; EGFR,
epidermal growth factor receptor; IRS-1, insulin receptor substrate-1;
PI3-kinase; phosphatidylinositol 3-kinase; SH2, Src homology 2;
CHO, Chinese hamster ovary; HA, hemagglutinin; NE,
norepinephrine; PY, anti-phosphotyrosine.
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REFERENCES |
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Top
Abstract
Introduction
Procedures
Results
Discussion
References
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- Leung, D. W., Spencer, S. A., Cachianes, G., Hammonds, R. G., Collins, C., Henzel, W. J., Barnard, R., Waters, M. J., and Wood, W. I. (1987) Nature 330, 537-543[CrossRef][Medline] [Order article via Infotrieve]
-
Cunningham, B. C.,
Ultsch, M.,
De Vos, A.,
Mulkerrin, M. G.,
Causer, K. R.,
and Wells, J. A.
(1991)
Science
254,
821-825
[Abstract/Free Full Text] - Argetsinger, L. S., Campbell, G. S., Yang, X., Witthuhn, B. A., Silvennoinen, O., Ihle, J. N., and Carter-Su, C. (1993) Cell 74, 237-244[CrossRef][Medline] [Order article via Infotrieve]
-
Rui, H.,
Kirken, R. A.,
and Farrar, W. L.
(1994)
J. Biol. Chem.
269,
5364-5368
[Abstract/Free Full Text] - O'Shea, J. J. (1997) Immunity 7, 1-11[CrossRef][Medline] [Order article via Infotrieve]
- Roupas, P., and Herington, A. C. (1994) Trends Endocrinol. Metab. 5, 154-158[CrossRef][Medline] [Order article via Infotrieve]
-
Winston, L. A.,
and Bertics, P. J.
(1992)
J. Biol. Chem.
267,
4747-4751
[Abstract/Free Full Text] -
Moller, C.,
Hanson, A.,
Enberg, B.,
Lobie, P. E.,
and Norstedt, G.
(1992)
J. Biol. Chem.
267,
23403-23408
[Abstract/Free Full Text] -
Peraldi, P.,
Scimeca, J.,
Filloux, C.,
and Van Obberghen, E.
(1993)
Endocrinology
132,
2578-2585
[Abstract/Free Full Text] - Rozakis-Adcock, M., McGlade, J., Mbamalu, G., Pelicii, G., Daly, R., Li, W., Batzer, A., Thomas, S., Brugge, J., Pelicci, P. G., Schlessinger, J., and Pawson, T. (1992) Nature 360, 689-692[CrossRef][Medline] [Order article via Infotrieve]
-
VanderKuur, J.,
Allevato, G.,
Billestrup, N.,
Norstedt, G.,
and Carter-Su, C.
(1995)
J. Biol. Chem.
270,
7587-7593
[Abstract/Free Full Text] - Yamauchi, T., Ueki, K., Tobe, K., Tamemoto, H., Sekine, N., Wada, M., Honjo, M., Takahashi, M., Takahashi, T., Hirai, H., Tushima, T., Akanuma, Y., Fujita, T., Komuro, I., Yazaki, Y., and Kadowaki, T. (1997) Nature 390, 91-96[CrossRef][Medline] [Order article via Infotrieve]
- Nicoll, C. S., Anderson, T. R., Hebert, N. J., and Russell, S. M. (1985) in Prolactin Basic and Clinical Correlates (MacLeod, R. M., Thorner, M. O., and Scapagnini, U., eds), pp. 393-413, Fidia Research Series, Liviana Press, Padua, Italy
-
Birnbaum, R. S.,
and Goodman, H. M.
(1976)
Endocrinology
99,
1336-1345
[Abstract/Free Full Text] - Honeyman, T. W., and Goodman, H. M. (1980) Am. J. Physiol. 238, 389-394
- Ridderstrale, M., and Tornqvist, H. (1994) Biochem. Biophys. Res. Commun. 203, 306-310[CrossRef][Medline] [Order article via Infotrieve]
- Sun, X. J., Rothenberg, P., Kahn, C. R., Backer, J. M., Araki, E., Wilden, P. A., Cahill, D. A., Goldstein, B. J., and White, M. F. (1991) Nature 352, 73-77[CrossRef][Medline] [Order article via Infotrieve]
-
White, M. F.,
and Kahn, C. R.
(1994)
J. Biol. Chem.
268,
1-4
[Abstract/Free Full Text] - Kadowaki, T., Tobe, K., Honda-Yamamoto, R., Tamemoto, H., Kaburagi, Y., Momomura, K., Ueki, K., Takahashi, Y., Yamauchi, T., Akanuma, Y., and Yazaki, Y. (1996) Endocr. J. 43, 33-41
- Backer, J. M., Myers, M. G., Jr., Shoelson, S. E., Chin, D. J., Sun, X. J., Miralpeix, M., Hu, P., Margolis, B., Skolnik, E. Y., Schlessinger, J., and White, M. F. (1992) EMBO J. 11, 3469-3479[Medline] [Order article via Infotrieve]
-
Lavan, B. E.,
Kuhne, M. R.,
Garner, C. W.,
Anderson, D.,
Reedijk, M.,
Pawson, T.,
and Lienhard, G. E.
(1992)
J. Biol. Chem.
267,
11631-11636
[Abstract/Free Full Text] -
Cheatham, B.,
Vlahos, C. J.,
Cheatham, L.,
Wang, L.,
Blenis, J.,
and Kahn, C. R.
(1994)
Mol. Cell. Biol.
14,
4902-4911
[Abstract/Free Full Text] -
Okada, T.,
Kawano, Y.,
Sakakibara, T.,
Hazeki, O.,
and Ui, M.
(1994)
J. Biol. Chem.
269,
3568-3573
[Abstract/Free Full Text] -
Yamamoto-Honda, R.,
Tobe, K.,
Kaburagi, Y.,
Ueki, K.,
Asai, S.,
Yachi, M.,
Shirouzu, M.,
Akanuma, Y.,
Yokoyama, S.,
Yazaki, Y.,
and Kadowaki, T.
(1995)
J. Biol. Chem.
270,
2729-2734
[Abstract/Free Full Text] - Araki, E., Lipes, M. A., Patti, M. E., Brüning, J. C., Haag, B., III., Johnson, R. S., and Kahn, C. R. (1994) Nature 372, 186-190[CrossRef][Medline] [Order article via Infotrieve]
- Tamemoto, H., Kadowaki, T., Tobe, K., Yagi, T., Sakura, H., Hayakawa, T., Terauchi, Y., Ueki, K., Kaburagi, Y., Satoh, S., Sekihara, H., Yoshioka, S., Horikoshi, H., Furuta, Y., Ikawa, Y., Kasuga, M., Yazaki, Y., and Aizawa, S. (1994) Nature 372, 182-186[CrossRef][Medline] [Order article via Infotrieve]
-
Wang, L-M.,
Myers, M. G., Jr.,
Sun, X-J.,
Aaronson, S. A.,
White, M. F.,
and Pierce, J. H.
(1993)
Science
261,
1591-1594
[Abstract/Free Full Text] - Sun, X. J., Wang, L. M., Zhang, Y., Yenush, L., Myers, M. G. J., Glasheen, E., Lane, W. S., Pierce, J. H., and White, M. F. (1995) Nature 377, 173-177[CrossRef][Medline] [Order article via Infotrieve]
-
Tobe, K.,
Tamemoto, H.,
Yamauchi, T.,
Aizawa, S.,
Yazaki, Y.,
and Kadowaki, T.
(1995)
J. Biol. Chem.
270,
5698-5701
[Abstract/Free Full Text] -
Patti, M.-E.,
Sun, X.-J.,
Bruening, J. C.,
Araki, E.,
Lipes, M. A.,
White, M. F.,
and Kahn, C. R.
(1995)
J. Biol. Chem.
270,
24670-24673
[Abstract/Free Full Text] - Yamauchi, T., Tobe, K., Tamemoto, H., Ueki, K., Kaburagi, Y., Yamamoto-Honda, R., Takahashi, Y., Yoshizawa, F., Aizawa, S., Akanuma, Y., Sonenberg, N., Yazaki, Y., and Kadowaki, T. (1996) Mol. Cell. Biol. 16, 3074-3084[Abstract]
- Momomura, K., Tobe, K., Seyama, Y., Takaku, F., and Kasuga, M. (1988) Biochem. Biophys. Res. Commun. 155, 1181-1186[CrossRef][Medline] [Order article via Infotrieve]
-
Lavan, B. E.,
Lane, W. S.,
and Lienhard, G. E.
(1997)
J. Biol. Chem.
272,
11439-11443
[Abstract/Free Full Text] -
Kaburagi, Y.,
Satoh, S.,
Tamemoto, H.,
Yamamoto-Honda, R.,
Tobe, K.,
Ueki, K.,
Yamauchi, T.,
Kono-Sugita, E.,
Sekihara, H.,
Aizawa, S.,
Cushman, S. W.,
Akanuma, Y.,
Yazaki, Y.,
and Kadowaki, T.
(1997)
J. Biol. Chem.
272,
25839-25844
[Abstract/Free Full Text] -
Souza, S. C.,
Frick, G. P.,
Yip, R.,
Lobo, R. B.,
Tai, L. R.,
and Goodman, H. M.
(1994)
J. Biol. Chem.
269,
30085-30088
[Abstract/Free Full Text] -
Ridderstrale, M.,
Degerman, E.,
and Tornqvist, H.
(1995)
J. Biol. Chem.
270,
3471-3474
[Abstract/Free Full Text] -
Argetsinger, L. S.,
Hsu, G. W.,
Myers, M. G., Jr.,
Billestrup, N.,
White, M. F.,
and Carter-Su, C.
(1995)
J. Biol. Chem.
270,
14685-14692
[Abstract/Free Full Text] -
Argetsinger, L. S.,
Norstedt, G.,
Billestrup, N.,
White, M. F.,
and Carter-Su, C.
(1996)
J. Biol. Chem.
271,
29415-29421
[Abstract/Free Full Text] -
Han, Y.,
Leaman, D. W.,
Watling, D.,
Rogers, N. C.,
Groner, B.,
Kerr, I. M.,
Wood, W. I.,
and Stark, G. R.
(1996)
J. Biol. Chem.
271,
5947-5952
[Abstract/Free Full Text] -
Watanabe, S.,
Itoh, T.,
and Arai, K.
(1996)
J. Biol. Chem.
271,
12681-12686
[Abstract/Free Full Text] -
Burfoot, M. S.,
Rogers, N. C.,
Watling, D.,
Smith, J. M.,
Pons, S.,
Paonessaw, G.,
Pellegrini, S.,
White, M. F.,
and Kerr, I. M.
(1997)
J. Biol. Chem.
272,
24183-24190
[Abstract/Free Full Text] -
Berlanga, J. J.,
Gualillo, O.,
Buteau, H.,
Applanat, M.,
Kelly, P. A.,
and Edery, M.
(1997)
J. Biol. Chem.
272,
2050-2052
[Abstract/Free Full Text] - Keegan, A. D., Nelms, K., White, M. F., Wang, L. M., Pierce, J. H., and Paul, W. E. (1994) Cell 76, 811-820[CrossRef][Medline] [Order article via Infotrieve]
-
O'Neill, T. J.,
Craparo, A.,
and Gustafson, T. A.
(1994)
Mol. Cell. Biol.
14,
6433-4772
[Abstract/Free Full Text] - Kaburagi, Y., Yamamoto-Honda, R., Tobe, K., Ueki, K., Yachi, M., Akanuma, Y., Stephens, D. K., Kaplan, D., Yazaki, Y., and Kadowaki, T. (1995) Endocrinology 136, 3437-3443[Abstract]
-
Mathews, L. S.,
Enberg, B.,
and Norstedt, G.
(1989)
J. Biol. Chem.
264,
9905-9910
[Abstract/Free Full Text] -
Silvennoinen, O.,
Witthuhn, B.,
Quelle, F. W.,
Cleveland, J. L.,
Yi, T.,
and Ihle, J. N.
(1993)
Proc. Natl. Acad. Sci. U. S. A.
90,
8429-8433
[Abstract/Free Full Text] -
Sakaue, H.,
Ogawa, W.,
Takata, M.,
Kuroda, S.,
Kotani, K.,
Matsumoto, M.,
Sakaue, M.,
Nishio, S.,
Ueno, H.,
and Kasuga, M.
(1997)
Mol. Endocrinol.
11,
1552-1562
[Abstract/Free Full Text]
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|
|
|
|
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|
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|
|
|
|
|
|
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|
|
|
|
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 K. Nishigaki, C. Hanson, T. Ohashi, D. Thompson, K. Muszynski, and S. Ruscetti Erythroid Cells Rendered Erythropoietin Independent by Infection with Friend Spleen Focus-Forming Virus Show Constitutive Activation of Phosphatidylinositol 3-Kinase and Akt Kinase: Involvement of Insulin Receptor Substrate-Related Adapter Proteins J. Virol., April 1, 2000; 74(7): 3037 - 3045. [Abstract] [Full Text]
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T. Zhu and P. E. Lobie Janus Kinase 2-dependent Activation of p38 Mitogen-activated Protein Kinase by Growth Hormone. RESULTANT TRANSCRIPTIONAL ACTIVATION OF ATF-2 AND CHOP, CYTOSKELETAL RE-ORGANIZATION AND MITOGENESIS J. Biol. Chem., January 21, 2000; 275(3): 2103 - 2114. [Abstract] [Full Text] [PDF]
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C. A. Peters, E. T. Maizels, and M. Hunzicker-Dunn Activation of PKC delta in the Rat Corpus Luteum during Pregnancy. POTENTIAL ROLE OF PROLACTIN SIGNALING J. Biol. Chem., December 24, 1999; 274(52): 37499 - 37505. [Abstract] [Full Text] [PDF]
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Y. Mao, P.-R. Ling, T. P. Fitzgibbons, K. C. McCowen, G. P. Frick, B. R. Bistrian, and R. J. Smith Endotoxin-Induced Inhibition of Growth Hormone Receptor Signaling in Rat Liver in Vivo Endocrinology, December 1, 1999; 140(12): 5505 - 5515. [Abstract] [Full Text]
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F. P. Dominici, D. Cifone, A. Bartke, and D. Turyn Alterations in the early steps of the insulin-signaling system in skeletal muscle of GH-transgenic mice Am J Physiol Endocrinol Metab, September 1, 1999; 277(3): E447 - E454. [Abstract] [Full Text] [PDF]
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T. Zhu, E. L. K. Goh, D. LeRoith, and P. E. Lobie Growth Hormone Stimulates the Formation of a Multiprotein Signaling Complex Involving p130Cas and CrkII. RESULTANT ACTIVATION OF c-Jun N-TERMINAL KINASE/STRESS-ACTIVATED PROTEIN KINASE (JNK/SAPK) J. Biol. Chem., December 11, 1998; 273(50): 33864 - 33875. [Abstract] [Full Text] [PDF]
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T. Yamauchi, N. Yamauchi, K. Ueki, T. Sugiyama, H. Waki, H. Miki, K. Tobe, S. Matsuda, T. Tsushima, T. Yamamoto, et al. Constitutive Tyrosine Phosphorylation of ErbB-2 via Jak2 by Autocrine Secretion of Prolactin in Human Breast Cancer J. Biol. Chem., October 20, 2000; 275(43): 33937 - 33944. [Abstract] [Full Text] [PDF]
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E. L. K. Goh, T. Zhu, S. Yakar, D. LeRoith, and P. E. Lobie CrkII Participation in the Cellular Effects of Growth Hormone and Insulin-like Growth Factor-1. PHOSPHATIDYLINOSITOL-3 KINASE DEPENDENT AND INDEPENDENT EFFECTS J. Biol. Chem., June 2, 2000; 275(23): 17683 - 17692. [Abstract] [Full Text] [PDF]
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