HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS		QUICK SEARCH:   [advanced] Author: Keyword(s): Year:  Vol:  Page:

This Article Abstract Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Citation Map Services Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Request Permissions Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Gadina, M. Articles by O’Shea, J. J. Search for Related Content PubMed PubMed Citation Articles by Gadina, M. Articles by O’Shea, J. J.

Cutting Edge: Involvement of SHP-2 in Multiple Aspects of IL-2 Signaling: Evidence for a Positive Regulatory Role

Massimo Gadina^1,*, Louis M. Stancato^*, Chris M. Bacon^2,*, Andrew C. Larner and John J. O’Shea^*

^* Lymphocyte Cell Biology Section, Arthritis and Rheumatism Branch, National Institute of Arthritis Musculoskeletal and Skin Diseases, National Institutes of Health, Bethesda, MD 20892; U.S. Food and Drug Administration, Center for Biologics Evaluation and Research, Division of Cytokine Biology, Bethesda, MD 20814

	Abstract

Top Abstract Introduction Materials and Methods Results and Discussion References

 
Binding of IL-2 to its receptor activates several biochemical pathways, but precisely how these pathways are linked is incompletely understood. Here, we report that SHP-2, an SH2-domain containing tyrosine phosphatase, associates with different molecules of the IL-2 signaling cascade. Upon IL-2 stimulation, SHP-2 was coimmunoprecipitated with Grb2 and the p85 subunit of phosphatidylinositol 3-kinase. In contrast, SHP-2 was constitutively associated with JAK1 and JAK3. Finally, SHP-2 expression amplified STAT-dependent transcriptional activation whereas a dominant negative allele inhibited transactivation and the IL-2-induced activation of MAPK (mitogen-activated protein kinase). These results demonstrate the involvement of SHP-2 in multiple pathways of the IL-2 signaling cascade and provide evidence for its positive regulatory role.

	Introduction

Top Abstract Introduction Materials and Methods Results and Discussion References

 
Interleukin 2 is a multifunctional cytokine that influences several cellular subsets of the immune system including T, B, and NK cells and monocytes (1). Binding of IL-2 to its receptor results in rapid tyrosine phosphorylation and activation of the Janus family protein-tyrosine kinases JAK1 and JAK3 (2, 3, 4). Substrates of the JAKs are the IL-2R itself (5), the STATs (6, 7, 8, 9), and the adapter molecule Shc (10, 11). The activation of Shc and subsequent binding to the adapter protein Grb2 is one means of activating the Ras/Raf/mitogen-activated protein kinase (MAPK)³ pathway. IL-2 also activates phosphatidylinositol 3' (PI 3)-kinase (12, 13) The mechanism by which it is activated has not been determined with certainty, although phosphorylation of insulin receptor substrate (IRS)-1 and IRS-2 provide one means of PI 3-kinase activation (14).

Precisely how these pathways are activated, how they are linked, and how they are regulated is incompletely understood. Many signaling pathways are regulated by the balance between tyrosine phosphorylation and dephosphorylation. Recently, many studies have demonstrated the importance of SHP-1 and SHP-2 (15), a subgroup of cytoplasmic protein-tyrosine phosphatases characterized by tandem phosphotyrosine-binding SH2 domains, in the regulation of signal transduction. SHP-1 is preferentially expressed in lymphohemopoietic cells and appears to act as a negative regulator of cytokine signaling, as demonstrated for IL-3, erythropoietin, and growth hormone (16, 17, 18). In contrast to SHP-1, SHP-2 appears to be much more widely expressed, but its exact function and substrates remain ill defined. It does not necessarily dephosphorylate receptors or kinases; indeed, some studies have demonstrated that it behaves as a positive regulator of cytokine signaling (19, 20, 21). In contrast, SHP-2 may inhibit TCR signaling (22).

Recently, it has been shown (23, 24) that SHP-2 is phosphorylated in response to IL-2 stimulation. However, the functions of SHP-2, either positive or negative, have not been characterized in IL-2 signaling. Here, we show that SHP-2 is involved in multiple pathways of IL-2 signaling, and we demonstrate its positive regulatory role.

	Materials and Methods

Top Abstract Introduction Materials and Methods Results and Discussion References

 
Immunoprecipitation and immunoblotting

Immunoprecipitations and immunoblottings were performed as previously described (5, 25) using commercially available Abs, with the exception of JAK3 (25) and SHP-2 and Tyk2 (21). Dephosphorylation experiments were performed by treating the lysates with 100 U of calf intestinal alkaline phosphatase at 37°C for 1 h before immunoprecipitation.

Cell culture and transfection

Human PBMC (97% CD3⁺) were prepared as described (26). NK3.3 and NIH3T3-ß cells (27) were kindly provided by Dr. J. Kornbluth, Arkansas Cancer Center, Little Rock, AR and Dr. T. Taniguchi, Tokyo University Faculty of Medicine, Tokyo, Japan, respectively. Before stimulation, cells were washed in CO₂-acidified RPMI and rested for 24 h.

Cells (2.8 x 10⁵) were transfected with 1 µg of p3xGAS-luciferase (provided by Dr. Richard Pine (Public Health Research Institute, New York, NY)) with the indicated amounts of pCMV5-WT-SHP-2, pCMV5-DN-SHP-2, pCMV5-SHP-1, or pCMV5-DN-SHP-1 (provided by Dr. Jack E. Dixon, University of Michigan Medical School, Ann Arbor, MI) and 0.8 µg of pCMV-ß (Clontech Laboratories, Palo Alto, CA) using Opti-MEM medium and Lipofectamine (Life Technologies, Gaithersburg, MD) for 8 h. After 16 h, the cells were stimulated with IL-2 (2000 IU/ml) for 8 h, and the luciferase and ß-gal activity was measured using the luciferase assay system (Promega, Madison, WI) and the galacto-light chemiluminescent reporter assays (Tropix, Bedford, MA), respectively.

MAPK assay

HA-ERK2 and pCMV5-DN-SHP-2 or pCMV5-DN-SHP-1 were transfected in NIH3T3-ß-JAK3 as described above. HA-MAPK was immunoprecipitated with an anti-hemagglutinin Ab, and used to phosphorylate myelin basic protein (MBP) (Sigma Chemical, St. Louis, MO). The reaction was conducted at room temperature for 15 min. SDS-PAGE and Western transfer were performed, and phosphorylated MBP was visualized and quantitated by Storm PhosphorImager (Molecular Dynamics, Sunnyvale, CA). The membrane was then immunoblotted using a monoclonal anti-pan MAPK Ab to confirm equivalent loading.

	Results and Discussion

Top Abstract Introduction Materials and Methods Results and Discussion References

 
IL-2-dependent binding of SHP-2 to Grb2 and PI 3-kinase

It has been previously shown that SHP-2 is phosphorylated in response to IL-2 (23, 24). IL-2 has been shown to activate MAPK, with two pathways contributing to this activation, the Grb2/SOS/Ras and the PI 3-kinase pathway (13). To clarify the function of SHP-2 in IL-2 signal transduction, we first analyzed potential interactions with Grb2 and PI 3-kinase. As shown in Figure 1, in the absence of stimulation, Grb2 and SHP-2 were not associated. However, following IL-2 stimulation, Grb2 associated with SHP-2 in NK3.3 cells (Fig. 1A, lanes 2 and 3) as well as in T cells (Fig. 1C, lane 2). Similarly, we observed IL-2-induced association with the p85 subunit of PI 3-kinase (Fig. 1, B, lanes 7–9, and D, lane 6). The specificity of the association was confirmed by immunoprecipitation with nonimmune serum and with anti-SHP-2 as a positive control. To confirm the SHP-2 associations, we also immunoprecipitated with anti-SHP-2 Ab and immunoblotted with anti p85 (Fig. 1E, lane 10). Similar results were obtained for Grb2 (data not shown).

View larger version (20K): [in this window] [in a new window]   FIGURE 1. IL-2 induces association of SHP-2 with Grb2 and p85 in NK and T cells. A, NK3.3 cells, untreated (lane 1) or IL-2 treated for 5 min (lanes 2, 4, and 5) or 20 min (lane 3), were lysed and immunoprecipitated with anti-Grb2 (lanes 1–3) or nonimmune serum (lane 4) (lane 5 represent a whole cells lysate) and then subjected to immunoblot analysis with anti-SHP-2 (top) or anti-Grb-2 (bottom). B, NK3.3 cells, untreated (lane 6) or IL-2 treated for 5 min (lane 7), 10 min (lane 8), or 15 min (lane 9), were lysed and immunoprecipitated with anti-p85 and then subjected to immunoblot analysis with anti-SHP-2 (top) or anti-p85 (bottom). C, T cells, untreated (lane 1) or IL-2 treated for 5 min (lanes 2–4), were lysed and immunoprecipitated with anti-Grb2 (lanes 1 and 2), anti-SHP-2 (lane 3) or nonimmune serum (lane 4) and then subjected to immunoblot analysis with anti-SHP-2 (top) or anti-Grb-2 (bottom). D, T cells, untreated (lane 5) or IL-2 treated for 5 min (lanes 6–8), were lysed and immunoprecipitated with anti-p85 (lanes 5 and 6), anti-SHP-2 (lane 7), or nonimmune serum (lane 8) and then subjected to immunoblot analysis with anti-SHP-2 (top) or anti-p85 (bottom). E, NK3.3 cells, untreated (lane 9 and 11) or IL-2 treated for 10 min (lanes 10 and 12), were lysed and immunoprecipitated with Santa Cruz anti-SHP-2 C18 (lanes 9 and 10) or nonimmune serum (lanes 11 and 12) and then subjected to immunoblot analysis with anti-p-85 (top) or anti-SHP-2 (bottom).

Previously, it has been shown that SHP-2 associates with JAKs, with the exception of JAK3, and may negatively regulate signaling (28). Therefore, we next studied the association of SHP-2 with the JAKs involved in IL-2 signaling. As shown in Figure 2, SHP-2 was associated with JAK3 (Fig. 2, A and C), JAK1 (B and D), and Tyk2 (not shown), but this association was not influenced by IL-2 treatment. Interestingly, the JAK-associated SHP-2 consistently migrated at a higher m.w. when compared with the majority of the directly immunoprecipitated SHP-2; note, however, that the total cellular SHP-2 detected by blotting lysates is more heterogeneous than the species that is/are immunoprecipitated (Fig. 2E, compare lane 3 with lane 4). The heterogeneity is likely due to phosphorylation on multiple serine, threonine, and tyrosine residues (29), and when lysates were subjected to partial in vitro dephosphorylation a portion of the JAK3-associated SHP-2 comigrated directly with the immunoprecipitated SHP-2 (Fig. 2, panel E, lane 2).

View larger version (18K): [in this window] [in a new window]   FIGURE 2. SHP-2 association with Janus kinases in NK and T cells. A, NK3.3 cells, untreated (lanes 1 and 3) or IL-2 treated for 5 min (lanes 2 and 4) or 15 min (lane 5), were lysed and immunoprecipitated with anti-SHP-2 (lanes 1 and 2) or anti-JAK3 (lanes 3–5), and then subjected to immunoblot analysis with anti-SHP-2 (top) or anti-JAK3 (bottom). B, NK3.3 cells, untreated (lane 6) or IL-2 treated for 5 min (lanes 7–9), were lysed and immunoprecipitated with anti-SHP-2 (lane 8), anti-JAK1 (lanes 6 and 7), or nonimmune serum (lane 9) and then subjected to immunoblot analysis with anti-SHP-2 (top) or anti-JAK1 (bottom). C, T cells, untreated (lanes 1 and 3) or IL-2 treated for 5 min (lanes 2, 4, and 5), were lysed and immunoprecipitated with anti-JAK3 (lanes 1 and 2), anti-SHP-2 (lanes 3 and 4), or nonimmune serum (lane 5) and then subjected to immunoblot analysis with anti-SHP-2 (top) or anti-JAK3 (bottom). D, T cells, untreated (lane 6) or IL-2 treated for 5 min (lanes 7 and 8), were lysed and immunoprecipitated with anti-JAK1 (lanes 6 and 7) or nonimmune serum (lane 8) and then subjected to immunoblot analysis with anti-SHP-2 (top) or anti-JAK1 (bottom). E, Lysates from NK3.3 cells were untreated (lanes 1 and 3) or treated with 100 U of alkaline phosphatase for 1 h (lanes 2 and 4), then immunoprecipitated with anti-JAK3 (lanes 1 and 2) or anti-SHP-2 (lane 3) (lane 4 represent a whole cells lysate). The immunoprecipitates were electophoresed, transferred, and then subjected to immunoblot analysis with anti-SHP-2 (top) and anti-JAK3 (bottom).

A previous report indicated that IL-2 induced SHP-2 phosphorylation was independent of JAK3 kinase activity (24). Therefore, we next examined whether the presence or absence of JAK3 affected the recruitment of SHP-2 to Grb2 and PI 3-kinase. We used NIH3T3 fibroblasts stably transfected with the IL-2R subunits with or without JAK3 (NIH3T3-ß; NIH3T3-ß-JAK3) (5, 27). Although in this immortalized factor-independent cell line, a basal level of association, not observed in T cells or in the NK3.3 cell line, was already present, we observed that association between SHP-2 and Grb2 was increased by IL-2 stimulation in cells expressing JAK3, whereas no increase was observed after IL-2 stimulation in cells lacking JAK3 (Fig. 3). Interestingly, IL-2-dependent PI 3-kinase/SHP-2 association was detectable in the absence of JAK3; however, it was clearly increased by the presence of JAK3. This association was less affected by the presence or absence of JAK3, suggesting a partially independently regulated signaling pathway. In NIH3T3-ß-JAK3, the basal level of PI 3-kinase/SHP-2 association is higher than in cells lacking JAK3, presumably because JAK3 is overexpressed in these cells.

View larger version (42K): [in this window] [in a new window]   FIGURE 3. IL-2-mediated SHP-2/Grb2 association is JAK3 dependent. A, Lysates from NIH3T3-ß and NIH3T3-ß-JAK3, untreated (lanes 1 and 3) or IL-2 treated for 5 min (lanes 2 and 4–6), were immunoprecipitated with anti-Grb2 (lanes 1–4), anti-SHP-2 (lane 5), or nonimmune serum (lane 6) and then subjected to immunoblot analysis with anti-SHP-2 (top) or anti-Grb-2 (bottom). B, Lysates from NIH3T3-ß and NIH3T3-ß-JAK3, untreated (lanes 7 and 9) or IL-2 treated for 5 min (lanes 8 and 10–12), were immunoprecipitated with anti-p85 (lanes 7–10), anti-SHP-2 (lane 11), or nonimmune serum (lane 12) and then subjected to immunoblot analysis with anti-SHP-2 (top) or anti-p85 (bottom).

Having demonstrated the association of SHP-2 with various molecules in the IL-2 signaling pathway, we next assessed its functional role. Specifically, in view of previous reports on experiments with other cytokines (28), we sought to determine whether SHP-2 plays a positive or a negative regulatory role in STAT activation. We therefore analyzed the effect of SHP-2 on IL-2-dependent transactivation using the IFN--activated sequence element of the IFN regulatory factor (IRF) gene fused to a luciferase reporter construct (30) (Fig. 4). The wild-type or catalytically inactive form of SHP-2 (DN-SHP-2) was cotransfected in NIH3T3-ß-JAK3 with the GAS luciferase construct, and the cells were stimulated. As shown in Figure 4A, IL-2 stimulation increased luciferase activity and addition of wild-type SHP-2 further enhanced this transactivation, whereas a related phosphatase, SHP-1, had relatively little effect. To further substantiate a positive regulatory role for SHP-2 in IL-2-dependent STAT activation, we next used expression of DN-SHP-2. As shown in Figure 4B, DN-SHP-2 significantly inhibited IL-2-mediated transactivation in a dose-dependent manner, whereas DN-SHP-1 had no effect. Equivalent transfection efficiency was confirmed by cotransfecting with a plasmid containing the ß-gal gene and measuring ß-gal activity on the transfectants.

View larger version (14K): [in this window] [in a new window]   FIGURE 4. SHP-2 but not SHP-1 positively regulates STAT-mediated transcriptional activation. NIH3T3-ß-JAK3 cells were transiently transfected with an IRF-reporter construct along with a control plasmid (E.V.), DN-SHP-2, wild-type SHP-2 (WT-SHP-2), DN-SHP-1, or WT-SHP-1 at the indicated concentrations for 8 h. The cells were placed in serum-free medium for 16 h, treated with (closed bars) or without (open bars) IL-2 (1000 IU/ml) for an additional 8 h, and then analyzed for luciferase activity. Results are expressed as luciferase activity (arbitrary light units) ± SD.

The positive regulatory role of SHP-2 in IL-2-dependent STAT activation is consistent with observations on another cytokine, prolactin (20); however, a mechanism is not immediately apparent. In contrast, Grb2 has clearly been demonstrated to be an intermediate that leads to MAPK activation. The IL-2-dependent association of SHP-2 with Grb2 suggested, therefore, that this phosphatase could play a role in the Grb2/SOS/Ras/MAPK pathway. We sought to verify this hypothesis by transfecting HA-tagged ERK2 with or without the DN-SHP-2. As shown in Figure 5, IL-2 dependent activation of MAPK was completely blocked by the overexpression of DN-SHP-2. To confirm the specificity of this effect, we also expressed (catalytically inactive) DN-SHP-1, which did not inhibit MAPK activation (data not shown). These findings also suggest that SHP-2 may have an essential role in coupling IL-2 signals to the activation of the MAPK pathway.

View larger version (40K): [in this window] [in a new window]   FIGURE 5. Catalytically inactive DN-SHP-2 inhibits MAPK activation. NIH3T3-ß-JAK3 cells were transiently transfected with an HA-tagged ERK2 construct along with an empty vector (E.V.) control and DN-SHP-2. Kinase assays were performed using MBP as a substrate. Incorporated radioactivity was analyzed using a Storm PhosphorImager (top), and the filter was subsequently immunoblotted with an anti-pan ERK Ab (bottom).

The regulation of signal transduction is maintained by a balance between positive and inhibitory signals. Previous reports suggested that SH2-containing phosphatases serve to dephosphorylate JAKs and, by inference, would be expected to blunt JAK-mediated signaling (17, 28). Our results, however, indicate that SHP-2 positively regulates STAT-dependent transcriptional activation, in accordance with studies on prolactin and IFN-ß signaling (20, 21). What is less clear is the mechanism by which SHP-2 might augment STAT-dependent transactivation. Our results suggest that by binding to Grb2 and PI 3-kinase, SHP-2 may exert a positive regulatory role in IL-2-induced stimulation of the MAPK pathway (13). Although SHP-2 might serve distinctive functions in the STAT, MAPK, and PI 3-kinase pathways, one may speculate that the action of SHP-2 on MAPK or PI 3-kinase could relate more directly to STAT activation. As STATs are also serine phosphorylated in response to cytokine activation (reviewed in 4 , this too may be a means by which SHP-2 could augment STAT-mediated transactivation. Whether the MAPK and STAT pathways intersect and whether SHP-2 functions in the manner outlined have yet to be ascertained. Nonetheless, these data suggest that SHP-2 may be involved in recruitment of essential molecules for the Ras/MAPK or the PI 3-kinase pathway. It will be important to better characterize these interactions and to dissect their respective functions. Although the target of SHP-2 catalytic activity has yet to be identified in any system, taken together, these findings suggest that SHP-2 may be important for regulating several of the IL-2-activated pathways.

	Acknowledgments

 
We thank Drs. J. Dixon and R. Pine for providing, respectively, SHP-2 and SHP-1 and IRF constructs; and Drs. J. Kornbluth and T. Taniguchi for providing, respectively, NK3.3 and NIH3T3-ß cells.

	Footnotes

 
¹ Address correspondence and reprint requests to Dr. Massimo Gadina, Building 10, Room 9N228, 10 Center Drive MSC 1820, Bethesda MD 20892-1820. E-mail address:

² C.M.B. was supported by a University of Sheffield Medical School PhD studentship, the Fullbright Commission, and the Yorkshire Cancer Research Campaign.

³ Abbreviations used in this paper: MAPK, mitogen-activated protein kinase; IRS, insulin receptor substrate; IRF, IFN regulatory factor; MBP, myelin basic protein; DN, dominant negative; PI 3, phosphatidylinositol 3'; HA, hemagglutinin.

Received for publication August 28, 1997. Accepted for publication March 20, 1998.

	References

Top Abstract Introduction Materials and Methods Results and Discussion References

 

1. Taniguchi, T., T. Miyazaki, Y. Minami, A. Kawahara, H. Fujii, Y. Nakagawa, M. Hatakeyama, Z. J. Liu. 1995. IL-2 signaling involves recruitment and activation of multiple protein tyrosine kinases by the IL-2 receptor. Ann. NY Acad. Sci. 766:235.[Medline]
2. Ihle, J. N.. 1996. Janus kinases in cytokine signaling. Philos. Trans. R. Soc. London B Biol. Sci. 351:159.[Medline]
3. Karnitz, L. M., R. T. Abraham. 1996. IL-2 receptor signaling mechanisms. Adv. Immunol. 61:147.[Medline]
4. O’Shea, J. J.. 1997. Jaks, STATs, cytokine signal transduction, and immunoregulation: are we there yet?. Immunity 7:1.[Medline]
5. Chen, M., A. Cheng, Y. Q. Chen, A. Hymel, E. P. Hanson, L. Kimmel, Y. Minami, T. Taniguchi, P. S. Changelian, J. J. O’Shea. 1997. The amino-termius of JAK3 is necessary and sufficient for binding to the common gamma chain and confers the ability to transmit interleukin 2-mediated signals. Proc. Natl. Acad. Sci. USA 94:6910.[Abstract/Free Full Text]
6. Fuji, H., Y. Nakagawa, U. Schindler, A. Kawahara, H. Mori, F. Gouilleux, B. Groner, J. N. Ihle, Y. Minami, T. Miyazaki, T. Taniguchi. 1995. Activation of Stat5 by IL-2 requires a carboxyl-terminal region of the IL-2 receptor ß chain but is not essential for the proliferative signal transmission. Proc. Natl. Acad. Sci. USA 92:5482.[Abstract/Free Full Text]
7. Hou, J., U. Schindler, W. J. Henzel, S. C. Wong, S. L. McKnight. 1995. Identification and purification of human Stat proteins activated in response to IL-2. Immunity 2:321.[Medline]
8. Johnston, J. A., C. M. Bacon, D. S. Finbloom, R. C. Rees, D. Kaplan, K. Shibuya, J. R. Ortaldo, S. Gupta, Y. Q. Chen, J. D. Giri, J.J. O’Shea. 1995. Tyrosine phosphorylation and activation of STAT5, STAT3, and Janus kinases by ILs 2 and 15. Proc. Natl. Acad. Sci. USA 92:8705.[Abstract/Free Full Text]
9. Lin, J. X., T. S. Migone, M. Tsang, M. Friedmann, J. A. Weatherbee, L. Zhou, A. Yamauchi, E. T. Bloom, J. Mietz, S. John, W. J. Leonard. 1995. The role of shared receptor motifs and common Stat proteins in the generation of cytokine pleiotropy and redundancy by IL-2, IL-4, IL-7, IL-13, and IL-15. Immunity 2:331.[Medline]
10. Burns, L. A., L. M. Karnitz, S. L. Sutor, R. T. Abraham. 1993. IL-2-induced tyrosine phosphorylation of p52shc in T lymphocytes. J. Biol. Chem. 268:17659.[Abstract/Free Full Text]
11. Ravichandran, K. S., S. J. Burakoff. 1994. The adapter protein Shc interacts with the interleukin-2 (IL-2) receptor upon IL-2 stimulation. J. Biol. Chem. 269:1599.[Abstract/Free Full Text]
12. Merida, I., E. Diez, G. N. Gaulton. 1991. IL-2 binding activates a tyrosine-phosphorylated phosphatidylinositol-3-kinase. J. Immunol. 147:2202.[Abstract]
13. Karnitz, L. M., L. A. Burns, S. L. Sutor, J. Blenis, R. T. Abraham. 1995. IL-2 triggers a novel phosphatidylinositol 3-kinase-dependent MEK activation pathway. Mol. Cell. Biol. 15:3049.[Abstract]
14. Johnston, J. A., L. M. Wang, E. P. Hanson, X. J. Sun, M. F. White, S. A. Oakes, J. H. Pierce, J. J. O’Shea. 1995. Interleukins 2, 4, 7, and 15 stimulate tyrosine phosphorylation of insulin receptor substrates 1 and 2 in T cells: potential role of JAK kinases. J. Biol. Chem. 270:28527.[Abstract/Free Full Text]
15. Thomas, M. L.. 1995. Positive and negative regulation of leukocyte activation by protein tyrosine phosphatases. Semin. Immunol. 7:279.[Medline]
16. Yi, T., A. L. Mui, G. Krystal, J. N. Ihle. 1993. Hematopoietic cell phosphatase associates with the interleukin-3 (IL-3) receptor beta chain and down-regulates IL-3-induced tyrosine phosphorylation and mitogenesis. Mol. Cell. Biol. 13:7577.[Abstract/Free Full Text]
17. Klingmuller, U., U. Lorenz, L. C. Cantley, B. G. Neel, H. F. Lodish. 1995. Specific recruitment of SH-PTP1 to the erythropoietin receptor causes inactivation of JAK2 and termination of proliferative signals. Cell 80:729.[Medline]
18. III. Petricoin, E., M. David, K. Igarashi, C. Benjamin, L. Ling, S. Goelz, D. S. Finbloom, A. C. Larner. 1996. Inhibition of alpha interferon but not gamma interferon signal transduction by phorbol esters is mediated by a tyrosine phosphatase. Mol. Cell. Biol. 16:1419.[Abstract]
19. Yamauchi, K., K. L. Milarski, A. R. Saltiel, J. E. Pessin. 1995. Protein-tyrosine-phosphatase SHPTP2 is a required positive effector for insulin downstream signaling. Proc. Natl. Acad. Sci. USA 92:664.[Abstract/Free Full Text]
20. Ali, S., Z. Chen, J. J. Lebrun, W. Vogel, A. Kharitonenkov, P. A. Kelly, A. Ullrich. 1996. PTP1D is a positive regulator of the prolactin signal leading to beta-casein promoter activation. EMBO J. 15:135.[Medline]
21. David, M., G. Zhou, R. Pine, J. E. Dixon, A. C. Larner. 1996. The SH2 domain-containing tyrosine phosphatase PTP1D is required for interferon alpha/beta-induced gene expression. J. Biol. Chem. 271:15862.[Abstract/Free Full Text]
22. Marengere, L. E., P. Waterhouse, G. S. Duncan, H. W. Mittrucker, G. S. Feng, T. W. Mak. 1996. Regulation of TCR signaling by tyrosine phosphatase SYP association with CTLA-4. Science 272:1170.[Abstract]
23. Nelson, B. H., B. C. McIntosh, L. L. Rosencrans, P. D. Greenberg. 1997. Requirement for an initial signal from the membrane-proximal region of the interleukin-2 receptor gamma(c) chain for Janus kinase activation leading to T cell proliferation. Proc. Natl. Acad. Sci. USA 94:1878.[Abstract/Free Full Text]
24. Adachi, M., M. Ishino, T. Torigoe, Y. Minami, T. Matozaki, T. Miyazaki, T. Taniguchi, Y. Hinoda, K. Imai. 1997. Interleukin-2 induces tyrosine phosphorylation of SHP-2 through IL-2 receptor beta chain. Oncogene 14:1629.[Medline]
25. Johnston, J. A., M. Kawamura, R. A. Kirken, Y. Q. Chen, T. B. Blake, K. Shibuya, J. R. Ortaldo, D. W. McVicar, J. J. O’Shea. 1994. Phosphorylation and activation of the Jak-3 Janus kinase in response to IL-2. Nature 370:151.[Medline]
26. Bacon, C. M., D. W. McVicar, J. R. Ortaldo, R. C. Rees, J. J. O’Shea, J. A. Johnston. 1995. Interleukin-12 (IL-12) induces tyrosine phosphorylation of JAK2 and TYK2: differential use of Janus family tyrosine kinases by IL-2 and IL-12. J. Exp. Med. 181:399.[Abstract/Free Full Text]
27. Minami, Y., I. Oishi, Z. J. Liu, S. Nakagawa, T. Miyazaki, T. Taniguchi. 1994. Signal transduction mediated by the reconstituted IL-2 receptor: evidence for a cell type-specific function of IL-2 receptor ß-chain. J. Immunol. 152:5680.[Abstract]
28. Sengupta, T. K., E. M. Schmitt, L. B. Ivashkiv. 1996. Inhibition of cytokines and JAK-STAT activation by distinct signaling pathways. Proc. Natl. Acad. Sci. USA 93:9499.[Abstract/Free Full Text]
29. Vogel, W., A. Ullrich. 1996. Multiple in vivo phosphorylated tyrosine phosphatase SHP-2 engages binding to Grb2 via tyrosine 584. Cell Growth Differ. 7:1589.[Abstract]
30. Pine, R. A., A. Canova, C. Schindler. 1994. Tyrosine phosphorylated p91 binds to a single element in the ISGF2/IRF1 promoter to mediate induction by IFN alpha and IFN gamma and is likely to autoregulate p91 gene. EMBO J. 13:158.[Medline]
31. Noguchi, T., T. Matozaki, K. Horita, Y. Fujioka, M. Kasuga. 1994. Role of SH-PTP2, a protein-tyrosine phosphatase with Src homology 2 domains, in insulin-stimulated Ras activation. Mol. Cell. Biol. 14:6674.[Abstract/Free Full Text]
32. Welham, M. J., U. Dechert, K. B. Leslie, F. Jirik, J. W. Schrader. 1994. Interleukin (IL)-3 and granulocyte/macrophage colony-stimulating factor, but not IL-4, induce tyrosine phosphorylation, activation, and association of SHPTP2 with Grb2 and phosphatidylinositol 3'-kinase. J. Biol. Chem. 269:23764.[Abstract/Free Full Text]

This article has been cited by other articles:

				T. Tomizawa, Y. Kaneko, Y. Kaneko, Y. Saito, H. Ohnishi, J. Okajo, C. Okuzawa, T. Ishikawa-Sekigami, Y. Murata, H. Okazawa, et al. Resistance to Experimental Autoimmune Encephalomyelitis and Impaired T Cell Priming by Dendritic Cells in Src Homology 2 Domain-Containing Protein Tyrosine Phosphatase Substrate-1 Mutant Mice J. Immunol., July 15, 2007; 179(2): 869 - 877. [Abstract] [Full Text] [PDF]

				R. J. Salmond, G. Huyer, A. Kotsoni, L. Clements, and D. R. Alexander The src Homology 2 Domain-Containing Tyrosine Phosphatase 2 Regulates Primary T-Dependent Immune Responses and Th Cell Differentiation J. Immunol., November 15, 2005; 175(10): 6498 - 6508. [Abstract] [Full Text] [PDF]

				M. Arnaud, C. Crouin, C. Deon, D. Loyaux, and J. Bertoglio Phosphorylation of Grb2-Associated Binder 2 on Serine 623 by ERK MAPK Regulates Its Association with the Phosphatase SHP-2 and Decreases STAT5 Activation J. Immunol., September 15, 2004; 173(6): 3962 - 3971. [Abstract] [Full Text] [PDF]

				N. Aoki, S. Ueno, H. Mano, S. Yamasaki, M. Shiota, H. Miyazaki, Y. Yamaguchi-Aoki, T. Matsuda, and A. Ullrich Mutual Regulation of Protein-tyrosine Phosphatase 20 and Protein-tyrosine Kinase Tec Activities by Tyrosine Phosphorylation and Dephosphorylation J. Biol. Chem., March 12, 2004; 279(11): 10765 - 10775. [Abstract] [Full Text] [PDF]

				N. Lerner-Marmarosh, M. Yoshizumi, W. Che, J. Surapisitchat, H. Kawakatsu, M. Akaike, B. Ding, Q. Huang, C. Yan, B. C. Berk, et al. Inhibition of Tumor Necrosis Factor-{alpha}-Induced SHP-2 Phosphatase Activity by Shear Stress: A Mechanism to Reduce Endothelial Inflammation Arterioscler. Thromb. Vasc. Biol., October 1, 2003; 23(10): 1775 - 1781. [Abstract] [Full Text] [PDF]

				V. Sriuranpong, J. I. Park, P. Amornphimoltham, V. Patel, B. D. Nelkin, and J. S. Gutkind Epidermal Growth Factor Receptor-independent Constitutive Activation of STAT3 in Head and Neck Squamous Cell Carcinoma Is Mediated by the Autocrine/Paracrine Stimulation of the Interleukin 6/gp130 Cytokine System Cancer Res., June 1, 2003; 63(11): 2948 - 2956. [Abstract] [Full Text] [PDF]

				S.-i. Yusa and K. S. Campbell Src Homology Region 2-Containing Protein Tyrosine Phosphatase-2 (SHP-2) Can Play a Direct Role in the Inhibitory Function of Killer Cell Ig-Like Receptors in Human NK Cells J. Immunol., May 1, 2003; 170(9): 4539 - 4547. [Abstract] [Full Text] [PDF]

				J. A. F. Vara, M. A. D. Caceres, A. Silva, and J. Martin-Perez Src Family Kinases Are Required for Prolactin Induction of Cell Proliferation Mol. Biol. Cell, July 1, 2001; 12(7): 2171 - 2183. [Abstract] [Full Text] [PDF]

				N. Fournier, L. Chalus, I. Durand, E. Garcia, J.-J. Pin, T. Churakova, S. Patel, C. Zlot, D. Gorman, S. Zurawski, et al. FDF03, a Novel Inhibitory Receptor of the Immunoglobulin Superfamily, Is Expressed by Human Dendritic and Myeloid Cells J. Immunol., August 1, 2000; 165(3): 1197 - 1209. [Abstract] [Full Text] [PDF]

				M. C. Bosco, R. E. Curiel, A. H. Zea, M. G. Malabarba, J. R. Ortaldo, and I. Espinoza-Delgado IL-2 Signaling in Human Monocytes Involves the Phosphorylation and Activation of p59hck 1 J. Immunol., May 1, 2000; 164(9): 4575 - 4585. [Abstract] [Full Text] [PDF]

				H. Huang and W. E. Paul Protein Tyrosine Phosphatase Activity Is Required for IL-4 Induction of IL-4 Receptor {alpha}-Chain J. Immunol., February 1, 2000; 164(3): 1211 - 1215. [Abstract] [Full Text] [PDF]

				C.-L. Yu, Y.-J. Jin, and S. J. Burakoff Cytosolic Tyrosine Dephosphorylation of STAT5. POTENTIAL ROLE OF SHP-2 IN STAT5 REGULATION J. Biol. Chem., January 7, 2000; 275(1): 599 - 604. [Abstract] [Full Text] [PDF]

				H. Lienard, P. Bruhns, O. Malbec, W. H. Fridman, and M. Daeron Signal Regulatory Proteins Negatively Regulate Immunoreceptor-dependent Cell Activation J. Biol. Chem., November 5, 1999; 274(45): 32493 - 32499. [Abstract] [Full Text] [PDF]

				P. Bruhns, P. Marchetti, W. H. Fridman, E. Vivier, and M. Daeron Differential Roles of N- and C-Terminal Immunoreceptor Tyrosine-Based Inhibition Motifs During Inhibition of Cell Activation by Killer Cell Inhibitory Receptors J. Immunol., March 15, 1999; 162(6): 3168 - 3175. [Abstract] [Full Text] [PDF]

				M. Gadina, C. Sudarshan, and J. J. O'Shea IL-2, But Not IL-4 and Other Cytokines, Induces Phosphorylation of a 98-kDa Protein Associated with SHP-2, Phosphatidylinositol 3'-Kinase, and Grb2 J. Immunol., February 15, 1999; 162(4): 2081 - 2086. [Abstract] [Full Text] [PDF]

				P. Bruhns, F. Vely, O. Malbec, W. H. Fridman, E. Vivier, and M. Daeron Molecular Basis of the Recruitment of the SH2 Domain-containing Inositol 5-Phosphatases SHIP1 and SHIP2 by Fcgamma RIIB J. Biol. Chem., November 22, 2000; 275(48): 37357 - 37364. [Abstract] [Full Text] [PDF]

				M. Gadina, C. Sudarshan, R. Visconti, Y.-J. Zhou, H. Gu, B. G. Neel, and J. J. O'Shea The Docking Molecule Gab2 Is Induced by Lymphocyte Activation and Is Involved in Signaling by Interleukin-2 and Interleukin-15 but Not Other Common gamma Chain-using Cytokines J. Biol. Chem., August 25, 2000; 275(35): 26959 - 26966. [Abstract] [Full Text] [PDF]

				N. Aoki and T. Matsuda A Cytosolic Protein-tyrosine Phosphatase PTP1B Specifically Dephosphorylates and Deactivates Prolactin-activated STAT5a and STAT5b J. Biol. Chem., December 8, 2000; 275(50): 39718 - 39726. [Abstract] [Full Text] [PDF]

				S. Ali and S. Ali Recruitment of the Protein-tyrosine Phosphatase SHP-2 to the C-terminal Tyrosine of the Prolactin Receptor and to the Adaptor Protein Gab2 J. Biol. Chem., December 8, 2000; 275(50): 39073 - 39080. [Abstract] [Full Text] [PDF]

This Article Abstract Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Citation Map Services Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Request Permissions Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Gadina, M. Articles by O’Shea, J. J. Search for Related Content PubMed PubMed Citation Articles by Gadina, M. Articles by O’Shea, J. J.