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(Received for publication, May 23, 1995) From the
Interleukin (IL)-9 stimulates the proliferation of a variety of
hematopoietic lineages through its interaction with a receptor of the
cytokine receptor superfamily. In the studies presented here, we have
begun to characterize the downstream signaling pathways activated by
IL-9. In addition to the activation of JAK1 and JAK3 tyrosine kinases,
IL-9, unlike most hematopoietic cytokines but similar to IL-4, induces
the tyrosine phosphorylation of a 170-kDa protein that is related to
the insulin receptor substrate-1 (IRS-1). We further demonstrate for
the first time that IRS-1 is not only associated with JAK1 but also
tyrosine phosphorylated and functionally involved in IL-9 signaling in
TS1 lymphocytes transfected with the murine IRS-1 cDNA. Cotransfection
studies and in vitro experiments directly demonstrate that
JAK1, JAK2, or JAK3 is capable of tyrosine phosphorylating IRS-1,
suggesting a functional role for these kinases in vivo.
Lastly, we demonstrate that IL-9 induces the tyrosine phosphorylation
of Stat3 and in this regard differs from IL-4, which triggers tyrosine
phosphorylation of Stat6. Taken together, these results strongly
suggest that IL-9 and IL-4 utilize common and unique signaling pathways
via inducing the similar and distinct tyrosine-phosphorylated proteins.
IL-9 ( IL-9 receptor belongs to the hematopoietin
receptor superfamily(10) . Although there is no intrinsic
tyrosine kinase motif in the IL-9 receptor, IL-9 has been shown to
stimulate protein tyrosine phosphorylation in D10G4.1 T
lymphocytes(11) . Recently, it has been demonstrated that JAK1
and JAK3 tyrosine kinases are activated following IL-9
stimulation(11, 12, 13) . Activation of JAK
tyrosine kinases has been linked to tyrosine phosphorylation and
activation of signal transducers and activators of transcription
(Stat)(14, 15) . However, it is not clear whether JAK
kinases can directly phosphorylate other proteins involved in IL-9
signaling. Here we show for the first time that insulin receptor
substrate-1 (IRS-1) and IRS-1-related IL-4-induced phosphorylated
substrate (4PS), which play essential roles in insulin- and
IL-4-mediated signal
transduction(16, 17, 18, 19) , are
not only rapidly tyrosine-phosphorylated but also functionally involved
in IL-9 signal transduction. Furthermore, we provide evidence that
activated JAK1, JAK2, and JAK3 tyrosine kinases induced by either IL-9
stimulation or overexpression in COS-1 cells can phosphorylate IRS-1 on
tyrosine residues both in vitro and in vivo. Our
results also indicate that IL-9 induces tyrosine phosphorylation of
Stat3, whereas IL-4 activates Stat6. These results strongly suggest
that IL-9 and IL-4 can trigger common and unique signaling pathways via
inducing the same and distinct tyrosine-phosphorylated proteins.
Figure 1:
Protein tyrosine
phosphorylation and activation of JAK1 and JAK3 kinases induced by IL-9
in TS1 cells. After factor starvation, TS1 cells (2
We also examined
whether IL-9 can induce tyrosine phosphorylation of known Stats. The
results in Fig. 1D demonstrated that Stat3 but not
Stat1, Stat2, Stat4, Stat5, or Stat6 is tyrosine-phosphorylated
following IL-9 stimulation in TS1 cells. In contrast, IL-4 specifically
induced Stat6 tyrosine phosphorylation in these cells (Fig. 1D). The results demonstrated that IL-4 and IL-9
utilize distinct Stats despite sharing IL-2 receptor
Figure 2:
Tyrosine phosphorylation of the IRS-1 and
IRS-1-related 4PS induced by IL-9 and IL-4 in TS1 cells. A,
tyrosine phosphorylation of IRS-1-related 4PS induced by IL-9 and IL-4.
Treatment and stimulation of cells with IL-9 and IL-4 were the same as
described in the legend to Fig. 1. Cell lysates (2
Because IRS-1-related
4PS cDNA and its specific antibody are not available at the present
time and IRS-1 and IRS-1-related 4PS share many biological
features(16, 18, 19) , we decided to
transfect IRS-1 cDNA into IRS-1-negative TS1 cells to further determine
whether IRS proteins are indeed involved in IL-9 signaling pathways.
The results in Fig. 2B showed that the overexpression
of IRS-1 in TS1 cells significantly enhances the sensitivity of TS1
cells to IL-9 and IL-4 stimulation (3-5-fold lower dosage
required to achieve the same level of cell proliferation when compared
with vector-transfected TS1 cells). Furthermore, we demonstrated that
transfected IRS-1 is rapidly tyrosine-phosphorylated and preferentially
associated with JAK1 kinase following IL-9 or IL-4 stimulation (Fig. 2C). These results strongly implicate that IRS-1
may be phosphorylated by JAK1 kinase and is functionally involved in
IL-9 signaling pathways in TS1 cells.
Figure 3:
Tyrosine phosphorylation of IRS-1 by JAK1,
JAK2, and JAK3 kinases. A and B, tyrosine
phosphorylation and association of IRS-1 with JAK1, JAK2, or JAK3
kinase in COS-1 cells. IRS-1 cDNA (4 µg) was cotransfected into
COS-1 cells with 6 µg of JAK1, JAK2, or JAK3 cDNAs as indicated. A, IRS-1 was immunoprecipitated with 4 µg of IRS-1
antibodies for 2 h at 4 °C after transfection for 48 h. B,
JAK1, JAK2 and JAK3 were immunoprecipitated from the same cell lysates
as in A with 2 µl of JAK1, JAK2, or JAK3 antisera as
indicated. The immunoprecipitates were separated by SDS-PAGE and
transferred to the PVDF membranes. The membranes were immunoblotted
with the indicated antibodies. C and D, in vitro tyrosine phosphorylation of IRS-1 by activated JAK1 and JAK3
kinases. The immunoprecipitated IRS-1 proteins from COS-1 cells
transfected with IRS-1 cDNA were mixed with the immunoprecipitated JAK1
or JAK3 kinase from either TS1 cells (C) stimulated without (JAK-TS1) or with IL-9 (JAK-TS1-IL-9) or COS-1 cells (D) transfected with JAK1 (JAK1-COS-1) or JAK3 cDNA (JAK3-COS-1) as indicated. The kinase reactions were initiated
by the addition of ATP as described under ``Materials and
Methods.'' The reaction mixtures were separated by SDS-PAGE (7.5%)
and transferred to the PVDF membranes. The membranes were immunoblotted
by ECL with anti-Tyr(P), anti-IRS-1, or anti-JAK1 and JAK3 antibodies
as indicated in the figure. PTyr,
Tyr(P).
We have shown that IL-9 and IL-4 induced common and unique
tyrosine-phosphorylated proteins in TS1 cells. IL-9 stimulated tyrosine
phosphorylation of Stat 3, whereas IL-4 activated Stat6. It is noted
that the cytoplasmic domain of IL-9 but not that of IL-2 or IL-4
receptor ligand binding subunit contains the motif YLPQ, which has been
shown to be a binding site for Stat3 in IL-6 signal transducer, gp130 (29) . These results suggest that unique motif(s) presented in
the cytoplasmic domain of cytokine receptors can determine specific
Stat activation. We previously showed that IL-9 but not IL-4 induces
tyrosine phosphorylation of Stat1 in D10G4.1 lymphocytes(11) .
In M07E megakaryocytic leukemia cells, Stat1 and 88-kDa associated
protein are activated by IL-9(12) . These results implicate
that activation of Stat by IL-9 may bear cell type specificity. Taken
together, although IL-4 and IL-9 share the IL-2 receptor It has
been demonstrated that the IRS-1 or IRS-1-related 4PS is essential for
signal transduction mediated by IL-4, insulin, and insulin-like growth
factor(27) . IRS-1 has also been shown to be involved in growth
hormone-mediated signaling(32) . We demonstrated here that
IRS-1 is not only tyrosine-phosphorylated but also functionally
involved in IL-9 signaling in TS1 cells. We further showed that IRS-1
is associated with JAK1 kinase following IL-9 or IL-4 stimulation,
implying that IRS-1 may be phosphorylated by JAK1 kinase in
vivo. It has been suggested that activation of JAK3 but not JAK1
may be critical in IL-4-mediated proliferative signaling(33) .
Although JAK3 is also activated by IL-9 in TS1 cells, we were unable to
show an association between JAK3 and IRS-1 using current extraction
procedures. It is possible that JAK3 may form a complex with IRS-1 in vivo; however, the association is disrupted during
extraction. Further studies using a two-hybrid system will be required
to verify such interactions. The conserved sequence motif
(PLX It has been shown that Stat proteins
are physiological substrates for JAK tyrosine kinases that can be
activated by a variety of cytokines (14, 15) . We
showed here that IRS-1 is tyrosine-phosphorylated and associated with
JAK1 in TS1 cells and both JAK1 and JAK2 are coimmunoprecipitated with
tyrosine-phosphorylated IRS-1 in COS-1 cotransfection experiments,
suggesting that IRS-1 is a substrate for JAK1 and JAK2 kinases.
Cotransfection of JAK3 with IRS-1 also resulted in tyrosine
phosphorylation of IRS-1, although we failed to show the direct
association between JAK3 and IRS-1. In vitro experiments
showed that activated JAK3 kinase is capable of phosphorylating IRS-1
on tyrosine residues, implying that IRS-1 is also a substrate for JAK3
kinase. It is well known that IRS-1 and IRS-1-related 4PS are
substrates for insulin and insulin-like growth factor receptor tyrosine
kinases and play essential roles in insulin and insulin-like growth
factor signaling
pathways(17, 18, 19, 34) . It has
been identified that the NPXY motif present in insulin
receptor cytoplasmic domain mediates interaction between IRS-1 and
insulin receptor tyrosine kinase(35) . Although the mechanisms
by which JAK kinases interact with IRS-1 require further investigation
because the NPXY motif is absent in JAK kinases, the
demonstration for the involvement of IRS-1 and IRS-1-related 4PS in
IL-9 signal transduction suggests that IL-9 shares some common
signaling pathways with those of IL-4, insulin, and insulin-like growth
factor. However, it remains to be answered whether JAK kinases and
insulin/insulin-like growth factor receptor tyrosine kinases can induce
the same sites of tyrosine phosphorylation in IRS-1 and IRS-1-related
4PS and whether the functions of tyrosine-phosphorylated IRS-1 and
IRS-1-related 4PS induced by these different tyrosine kinases are the
same in vivo.
Volume 270,
Number 35,
Issue of September 01, pp. 20497-20502, 1995
©1995 by The American Society for Biochemistry and Molecular Biology, Inc.
ABSTRACT
INTRODUCTION
MATERIALS AND METHODS
RESULTS
DISCUSSION
FOOTNOTES
ACKNOWLEDGEMENTS
REFERENCES
)is a T cell-derived lymphokine with growth
promoting activity for certain murine T helper clones(1) . The
cDNAs encoding murine and human IL-9 have been
cloned(2, 3) . Subsequently, it was found that IL-9
has a variety of biological activities including T cell and mast cell
growth-promoting activity(4, 5) , B cell
differentiation activity(6) , and stimulatory activity toward
erythroid and myeloid precursors(7, 8) . Recently, the
possible involvement of IL-9 in T cell tumorigenesis has been
suggested(9) . These results suggest that IL-9 is a
multifunctional T cell cytokine that may play an important role in
immunohematopoiesis.
Reagents and Cytokines
Antibodies for JAK1,
JAK2, JAK3, TyK2, Stat2, IRS-1 (polyclonal antibodies against the
intact molecule and the C terminus of the protein), and phosphotyrosine
(Tyr(P), 4G10) were purchased from Upstate Biotechnology Inc. (Lake
Placid, NY). Antibodies for the N terminus of IRS-1 and Stat1 were
purchased from Santa Cruz (Santa Cruz, CA). Antibodies for Stat3,
Stat4, Stat5, and Stat6 were produced as described(20) . ECL
detection kit was purchased from Amersham Corp. Recombinant murine IL-9
and IL-4 were from R & D Systems (Minneapolis, MN). DOTAP
transfection reagents were purchased from Boehringer Mannheim.Cell Culture and Cytokine
Stimulation
IL-9-dependent murine T cell line (TS1) was
maintained in Click's medium (Irvine Scientific) plus 10% fetal
bovine serum in the presence of 0.1 ng/ml murine IL-9(1) . For
experiments, TS1 cells were washed three times with serum-free medium,
starved for 7 h in the absence of IL-9, and washed once every 2 h with
serum-free medium during starvation. Cells (2 10
cells/ml) were stimulated with IL-9 (50 ng/ml) or IL-4 (50 ng/ml)
for the indicated periods of time and lysed with 1% Nonidet P-40 lysis
buffer as described(21) .Immunoprecipitation and Immunoblotting
Anti-JAK1,
JAK2, JAK3, TyK2, Stat, and IRS-1 immunoprecipitations were done as
described (20, 21, 22) with minor
modifications (50 mM Tris-HCl, pH 7.4, 150 mM NaCl,
0.2 mM EDTA, and 0.5% Nonidet P-40 lysis buffer for
coimmunoprecipitation). Briefly, cell lysates (2 10
cells/sample) were incubated with 1 (JAK1, JAK2, JAK3, or TyK2)
or 2 µl (Stat3, Stat4, Stat5, or Stat6) of antisera and 1 µg of
Stat1, 20 µg of Stat2, or 3 µg of IRS-1 (C terminus) antibodies
for 2 h at 4 °C or 10 µg of anti-IRS-1 antibodies against the
intact IRS-1 overnight at 4 °C with rotation, and then protein
A-agarose beads were added for an additional 1 h. The precipitates were
washed five times with lysis buffer and proteins were dissociated from
protein A-agarose beads by boiling for 5 min with SDS-PAGE sample
buffer. Proteins were separated by SDS-PAGE (7.5%) and transferred to
polyvinylidene difluoride (PVDF) membranes. The membranes were
immunoblotted with antibodies for Tyr(P) (0.1 µg/ml), JAK1 (1:500),
JAK2 (1:1000), JAK3 (1:1000), Stat1 (2 µg/ml), Stat2 (5 µg),
Stat3/Stat4/Stat5/Stat6 (1:2000), or IRS-1 (2 µg/ml for the C
terminus and 10 µg/ml for the intact molecule) as indicated under
``Results'' and ``Discussion.''Expression Vectors and Transfections
The
cDNA-encoding murine IRS-1(23) , JAK1, JAK2, and JAK3 (24, 25, 26) kinases were subcloned into the
mammalian expression vectors pRc/CMV (Invitrogen). TS1 cells were
transfected with IRS-1 cDNA or vector alone by electroporation and
selected with G418 (1 mg/ml). The G418-resistant cells were further
cloned by limiting dilution, and 40 clones were picked for screening
IRS-1 expression by immunoblotting. Two clones that expressed high
levels of IRS-1 were used for experiments. For cotransfection,
subconfluent COS-1 cells in 100-mm culture dishes were transfected with
various combinations of IRS-1, JAK1, JAK2, or JAK3 expression
constructs by the DOTAP method according to the manufacturer's
instructions. Cells were lysed with lysis buffer as described above
following transfection for 48 h, and proteins were immunoprecipitated
with the indicated antibodies.In Vitro Kinase Assay
Anti-JAK1 and JAK3 antibody
immunoprecipitates were washed five times with lysis buffer and once
with kinase buffer. The in vitro kinase assay was performed
and visualized as described(21, 22) . For in vitro tyrosine phosphorylation of IRS-1, JAK1 or JAK3 kinase was
immunoprecipitated and washed with kinase buffer minus ATP and then
mixed with immunoprecipitated IRS-1. The kinase reactions were
initiated by adding 100 µM of ATP and incubated for 30 min
at 30 °C as described previously (21, 22, 27) and indicated under
``Results.''
Characterization of Tyrosine-phosphorylated Proteins
Induced by IL-9 in TS1 Lymphocytes
As shown in Fig. 1A, IL-9 induced multiple tyrosine-phosphorylated
proteins with apparent molecular masses of 170, 145, 130, 125, 97, 89,
67, 64, and 40 kDa in TS1 cells. We have identified the 170-, 130-, and
125-kDa tyrosine-phosphorylated proteins as the IRS-1-related 4PS
protein, JAK1, and JAK3 tyrosine kinases, respectively (Fig. 1, B and C, and 2A). Furthermore, JAK1 and JAK3
kinases were activated according to in vitro autophosphorylation reactions following IL-9 stimulation (Fig. 1C). JAK2 and TyK2 kinases, however, were not
activated by IL-9 in TS1 cells (data not shown). The 89-kDa
tyrosine-phosphorylated protein is likely to be Stat3 (see below). The
40-kDa tyrosine-phosphorylated protein is unlikely to be
mitogen-activated protein kinases, because its molecular size is
different from those of mitogen-activated protein kinases and IL-9 did
not induce mitogen-activated protein kinase tyrosine phosphorylation in
TS1 cells (data not shown), suggesting that IL-9, similar to IL-4 but
unlike most cytokines, may not utilize mitogen-activated protein kinase
signaling cascades. The identity of the 67- and 64-kDa
tyrosine-phosphorylated proteins requires further investigation. In
comparison with IL-9, IL-4 induced the common (170-, 145-, 130-, 125-,
67-, and 64-kDa) and distinct (86- and 72-kDa) tyrosine-phosphorylated
proteins in TS1 cells (Fig. 1A).
10
cells in 0.5 ml of serum-free medium) were stimulated with 50
ng/ml of IL-9 or IL-4 for 5 min (A and D) or for the
indicated periods of time (B and C). Cells were lysed
with 1% Nonidet P-40 lysis buffer. A, tyrosine phosphorylation
of cellular proteins induced by IL-9 and IL-4. Equal amounts of total
cell lysates (4 10
cells/lane) were separated by
SDS-PAGE (7.5%), and proteins were transferred to PVDF membranes. B, kinetics of tyrosine phosphorylation of JAK1 and JAK3
kinases induced by IL-9. Cell lysates (2 10
cells/sample) were incubated with 1 µl of JAK1 or JAK3
antisera for 2 h at 4 °C. The immune complexes were precipitated by
the addition of protein A-agarose beads and washed with lysis buffer as
indicated under ``Materials and Methods.'' The
immunoprecipitates were separated by SDS-PAGE (7.5%) and transferred to
PVDF membranes. The membranes from A and B were
immunoblotted with the indicated antibodies. C, JAK1 and JAK3 in vitro kinase activity induced by IL-9. The
immunoprecipitates with JAK1 or JAK3 antisera as described in B were washed five times with lysis buffer and once with kinase
buffer. The immune complex kinase assays were performed by adding
[-
P]ATP as described under ``Materials
and Methods.'' The products of kinase reactions were resolved by
SDS-PAGE (7.5%) and transferred to PVDF membranes. The kinase activity
was detected by autoradiography, and the membranes were then
immunoblotted with JAK1 or JAK3 antisera as indicated. D,
tyrosine phosphorylation of Stat proteins induced by IL-9 and IL-4. The
Stat proteins were immunoprecipitated with specific Stat antibodies as
indicated and separated by SDS-PAGE (7%). The proteins were then
transferred to PVDF membranes and immunoblotted with the indicated
antibodies. PTyr, Tyr(P). IP,
immunoprecipitation.
chain and
activating the same JAK tyrosine kinases(11, 13) ,
implying that signaling specificity for each cytokine may be achieved
by activating distinct Stat.
Identification of Tyrosine Phosphorylation of the IRS-1
and IRS-1-related 4PS Induced by IL-9 in TS1 Cells
As shown in Fig. 1A, IL-9 induced tyrosine phosphorylation of a
170-kDa protein. IL-4 also induced tyrosine phosphorylation of a
170-kDa protein in TS1 cells. It has been shown that the 170-kDa
tyrosine-phosphorylated protein induced by IL-4 in a variety of cell
lines is the IRS-1 related 4PS
protein(16, 18, 19) . To test whether the
170-kDa tyrosine-phosphorylated protein in IL-9 signaling is 4PS,
immunoprecipitation experiments with anti-intact IRS-1 antibodies were
performed. The results demonstrated that the 170-kDa
tyrosine-phosphorylated protein induced by IL-9 or IL-4 in TS1 cells
can be precipitated with antibodies for intact IRS-1, although
efficiency seems low when compared with total cell lysates (Fig. 1A and Fig. 2A). Normal rabbit
IgG did not precipitate the 170-kDa tyrosine-phosphorylated protein
(data not shown). Furthermore, antibodies against the C or the N
terminus of the IRS-1 protein did not immunoprecipitate the 170-kDa
tyrosine-phosphorylated protein induced by IL-9 or IL-4 in TS1 cells
(data not shown). These results suggest that TS1 cells do not express
IRS-1 and that the 170-kDa tyrosine-phosphorylated protein induced by
IL-9 or IL-4 in TS1 cells shares some antigenic epitopes with IRS-1.
Based on the characteristics of this protein, it is most likely that
the 170-kDa tyrosine-phosphorylated protein induced by IL-9 in TS1
cells is the IRS-1-related 4PS(19) .
10
cells/sample) were incubated with 10 µg of
anti-IRS-1 antibodies raised against the intact IRS-1 protein overnight
at 4 °C. B, comparison of cell proliferation in TS1 cells
transfected with vector alone (TS1) or with murine IRS-1 cDNA (TS1-IRS-1) following IL-9 or IL-4 stimulation. Cells (200
cells/0.2 ml/well in triplicate) were incubated in 96-well plates for 3
days with different concentrations of IL-9 or IL-4 as indicated. The
number of cells in each well was counted by cell counter at the end of
culture. The results represent one of three different individual
experiments. C, association of IRS-1 with JAK1 kinase. TS1
cells transfected with IRS-1 cDNA were starved and stimulated with IL-9
or IL-4 as described in the legend to Fig. 1. Cells (2
10
cells/sample) were lysed, and IRS-1 was
immunoprecipitated with 3 µg of antibodies for the C terminus of
IRS-1 for 2 h at 4 °C. The immune complexes from A and C were precipitated by the addition of protein A-agarose beads
as described under ``Materials and Methods.'' The
immunoprecipitates were separated by SDS-PAGE (7.5%) and transferred to
PVDF membranes. The membranes were sequentially immunoblotted with
antibodies for Tyr(P) (PTyr), intact IRS-1 (A), or C
terminus of IRS-1 (C) and JAK1 or JAK3 (C) as
indicated. IP,
immunoprecipitation.
Demonstration for Tyrosine Phosphorylation of IRS-1 by
JAK Tyrosine Kinases
We have demonstrated that IL-9 induces
tyrosine phosphorylation of IRS-1 and IRS-1-related 4PS in TS1 T cells.
It is important to identify possible tyrosine kinases responsible for
tyrosine phosphorylation of IRS-1 and IRS-1-related 4PS in IL-9
signaling pathways. Because IL-9 activates JAK1 and JAK3 tyrosine
kinases, we hypothesize that JAK1 and/or JAK3 may be tyrosine kinases
capable of phosphorylating IRS-1 and IRS-1-related 4PS. It is known
that overexpression of JAK kinases resulted in kinase
autophosphorylation and activation (28) and that cotransfection
of JAK kinase expression constructs with plasmids containing Stat cDNAs
can result in tyrosine phosphorylation and activation of
Stats(14, 20, 28) . To directly test our
hypothesis, we subcloned IRS-1, JAK1, JAK2, and JAK3 cDNAs into
mammalian cell expression vector pRc/CMV and performed cotransfection
experiments with various combinations of IRS-1, JAK1, JAK2, or JAK3
cDNA plasmids in COS-1 cells. The results in Fig. 3, A and B, clearly indicated that cotransfection of IRS-1
plasmid with JAK1, JAK2, or JAK3 plasmids resulted in not only tyrosine
phosphorylation and activation of JAK kinases but also tyrosine
phosphorylation of IRS-1. Fig. 3A shows that JAK1 and
JAK2 are specifically coimmunoprecipitated with IRS-1 because
anti-IRS-1 antibodies did not precipitate any JAK kinases in COS-1
cells transfected with JAKs alone. (
)The association of
IRS-1 with JAK3 is too weak to be detected in these experiments. The
efficiency of coprecipitation of IRS-1 with anti-JAK antibodies seems
extremely low and is under detection (data not shown). The results from
COS-1 cell transfection experiments strongly implicate that IRS-1 is a
substrate for JAK kinases. Furthermore, in vitro experiments
with immunoprecipitated JAK1 or JAK3 kinase activated by IL-9 in TS1
cells or overexpression in COS-1 cells showed that activated JAK
kinases can phosphorylate IRS-1 on tyrosine residues (Fig. 3, C and D). Based on our cotransfection experiments in
COS-1 cells and in vitro experiments, we conclude that JAK1,
JAK2, and JAK3 kinases can directly interact with IRS-1 both in
vitro and in vivo and that JAK 1 and JAK3 kinases are
responsible for tyrosine phosphorylation of IRS-1 in IL-9 and IL-4
signaling pathways in T cells.
chain(13) , induce tyrosine phosphorylation of IRS-1/4PS, and
activate the same JAK1 and JAK3 kinases, the activation of distinct
Stat and other tyrosine-phosphorylated substrates suggests that the
specificity for IL-9 and IL-4 signaling could be achieved through the
unique tyrosine-phosphorylated proteins. For example, dysregulated
expression of IL-9 but not IL-4 can induce T cell
transformation(29, 30) , and IL-9 has erythroid
progenitor stimulating activity(8) , whereas IL-4 lacks this
function. On the other hand, the common signaling pathways triggered by
IL-4 and IL-9 may explain some of the overlapping activities of IL-4
and IL-9 such as stimulation of T cell proliferation as well as
induction of B cell differentiation(5, 31) .
NPXYXSXSD) (27) , which interacts with IRS-1 or IRS-1-related 4PS in IL-4,
insulin, and insulin-like growth factor receptors, is not present in
the IL-9 receptor(10) . Therefore, the mechanisms by which IL-9
induces tyrosine phosphorylation of the IRS-1 and IRS-1-related 4PS
require further investigation.
))
We thank Dr. J. Van Snick at the Ludwig Institute for
Cancer Research for providing the TS1 lymphocytes.
©1995 by The American Society for Biochemistry and Molecular Biology, Inc.
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 H. Ueno, E. Kondo, R. Yamamoto-Honda, K. Tobe, T. Nakamoto, K. Sasaki, K. Mitani, A. Furusaka, T. Tanaka, Y. Tsujimoto, et al. Association of Insulin Receptor Substrate Proteins with Bcl-2 and Their Effects on Its Phosphorylation and Antiapoptotic Function Mol. Biol. Cell, February 1, 2000; 11(2): 735 - 746. [Abstract] [Full Text]
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A. Wickrema, S. Uddin, A. Sharma, F. Chen, Y. Alsayed, S. Ahmad, S. T. Sawyer, G. Krystal, T. Yi, K. Nishada, et al. Engagement of Gab1 and Gab2 in Erythropoietin Signaling J. Biol. Chem., August 27, 1999; 274(35): 24469 - 24474. [Abstract] [Full Text] [PDF]
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 H. B. Sun, Y. X. Zhu, T. Yin, G. Sledge, and Y.-C. Yang MRG1, the product of a melanocyte-specific gene related gene, is a cytokine-inducible transcription factor with transformation activity PNAS, November 10, 1998; 95(23): 13555 - 13560. [Abstract] [Full Text] [PDF]
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L. Grasso, M. Huang, C. D. Sullivan, C. J. Messler, M. B. Kiser, C. R. Dragwa, K. J. Holroyd, J.-C. Renauld, R. C. Levitt, and N. C. Nicolaides Molecular Analysis of Human Interleukin-9 Receptor Transcripts in Peripheral Blood Mononuclear Cells. IDENTIFICATION OF A SPLICE VARIANT ENCODING FOR A NONFUNCTIONAL CELL SURFACE RECEPTOR J. Biol. Chem., September 11, 1998; 273(37): 24016 - 24024. [Abstract] [Full Text] [PDF]
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P. A. Goodman, L. B. Niehoff, and F. M. Uckun Role of Tyrosine Kinases in Induction of the c-jun Proto-oncogene in Irradiated B-lineage Lymphoid Cells J. Biol. Chem., July 10, 1998; 273(28): 17742 - 17748. [Abstract] [Full Text] [PDF]
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S. Skov, M. Nielsen, S. Bregenholt, N. Odum, and M. H. Claesson Activation of Stat-3 Is Involved in the Induction of Apoptosis After Ligation of Major Histocompatibility Complex Class I Molecules on Human Jurkat T Cells Blood, May 15, 1998; 91(10): 3566 - 3573. [Abstract] [Full Text] [PDF]
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R. G. Richards, M. P. Walker, J. Sebastian, and R. P. DiAugustine Insulin-like Growth Factor-1 (IGF-1) Receptor-Insulin Receptor Substrate Complexes in the Uterus. ALTERED SIGNALING RESPONSE TO ESTRADIOL IN THE IGF-1m/m MOUSE J. Biol. Chem., May 8, 1998; 273(19): 11962 - 11969. [Abstract] [Full Text] [PDF]
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J. H. Bauer, K. D. Liu, Y. You, S. Y. Lai, and M. A. Goldsmith Heteromerization of the gamma c Chain with the Interleukin-9 Receptor alpha Subunit Leads to STAT Activation and Prevention of Apoptosis J. Biol. Chem., April 10, 1998; 273(15): 9255 - 9260. [Abstract] [Full Text] [PDF]
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D. Guo, J. D. Dunbar, C. H. Yang, L. M. Pfeffer, and D. B. Donner Induction of Jak/STAT Signaling by Activation of the Type 1 TNF Receptor J. Immunol., March 15, 1998; 160(6): 2742 - 2750. [Abstract] [Full Text] [PDF]
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 P. Gual, V. Baron, V. Lequoy, and E. Van Obberghen Interaction of Janus Kinases JAK-1 and JAK-2 with the Insulin Receptor and the Insulin-Like Growth Factor-1 Receptor Endocrinology, March 1, 1998; 139(3): 884 - 893. [Abstract] [Full Text] [PDF]
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S. Uddin, E. N. Fish, D. Sher, C. Gardziola, O. R. Colamonici, M. Kellum, P. M. Pitha, M. F. White, and L. C. Platanias The IRS-Pathway Operates Distinctively From the Stat-Pathway in Hematopoietic Cells and Transduces Common and Distinct Signals During Engagement of the Insulin or Interferon-alpha Receptors Blood, October 1, 1997; 90(7): 2574 - 2582. [Abstract] [Full Text] [PDF]
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Y. X. Zhu, H. B. Sun, M. L.-S. Tsang, J. McMahel, S. Grigsby, T. Yin, and Y.-C. Yang Critical Cytoplasmic Domains of Human Interleukin-9 Receptor alpha Chain in Interleukin-9-mediated Cell Proliferation and Signal Transduction J. Biol. Chem., August 22, 1997; 272(34): 21334 - 21340. [Abstract] [Full Text] [PDF]
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T. Yin, G. Sandhu, C. D. Wolfgang, A. Burrier, R. L. Webb, D. F. Rigel, T. Hai, and J. Whelan Tissue-specific Pattern of Stress Kinase Activation in Ischemic/Reperfused Heart and Kidney J. Biol. Chem., August 8, 1997; 272(32): 19943 - 19950. [Abstract] [Full Text] [PDF]
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H. Fujiwara, S. H. Hanissian, A. Tsytsykova, and R. S. Geha Homodimerization of the human interleukin 4 receptor alpha chain induces Cvarepsilon germline transcripts in B cells in the absence of the interleukin 2 receptor gamma chain PNAS, May 27, 1997; 94(11): 5866 - 5871. [Abstract] [Full Text] [PDF]
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X. H. Chen, B. K.R. Patel, L.-M. Wang, M. Frankel, N. Ellmore, R. A. Flavell, W. J. LaRochelle, and J. H. Pierce Jak1 Expression Is Required for Mediating Interleukin-4-induced Tyrosine Phosphorylation of Insulin Receptor Substrate and Stat6 Signaling Molecules J. Biol. Chem., March 7, 1997; 272(10): 6556 - 6560. [Abstract] [Full Text] [PDF]
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J. J. Berlanga, O. Gualillo, H. Buteau, M. Applanat, P. A. Kelly, and M. Edery Prolactin Activates Tyrosyl Phosphorylation of Insulin Receptor Substrate 1and Phosphatidylinositol-3-OH Kinase J. Biol. Chem., January 24, 1997; 272(4): 2050 - 2052. [Abstract] [Full Text] [PDF]
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T. Yin, R. Shen, G.-S. Feng, and Y.-C. Yang Molecular Characterization of Specific Interactions between SHP-2 Phosphatase and JAK Tyrosine Kinases J. Biol. Chem., January 10, 1997; 272(2): 1032 - 1037. [Abstract] [Full Text] [PDF]
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H. Ueno, K. Sasaki, H. Kozutsumi, K. Miyagawa, K. Mitani, Y. Yazaki, and H. Hirai Growth and Survival Signals transmitted via Two Distinct NPXY Motifs within Leukocyte Tyrosine Kinase, an Insulin Receptor-related Tyrosine Kinase J. Biol. Chem., November 1, 1996; 271(44): 27707 - 27714. [Abstract] [Full Text] [PDF]
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A. Kowalski-Chauvel, L. Pradayrol, N. Vaysse, and C. Seva Gastrin Stimulates Tyrosine Phosphorylation of Insulin Receptor Substrate 1and Its Association with Grb2 and the Phosphatidylinositol 3-Kinase J. Biol. Chem., October 18, 1996; 271(42): 26356 - 26361. [Abstract] [Full Text] [PDF]
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L. C. Platanias, S. Uddin, A. Yetter, X.-J. Sun, and M. F. White The Type I Interferon Receptor Mediates Tyrosine Phosphorylation of Insulin Receptor Substrate 2 J. Biol. Chem., January 5, 1996; 271(1): 278 - 282. [Abstract] [Full Text] [PDF]
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