Presentation is loading. Please wait.

Presentation is loading. Please wait.

Roles of JAK kinases in human GM-CSF receptor signal transduction

Published byJacob Walton Modified over 5 years ago

Similar presentations

Presentation on theme: "Roles of JAK kinases in human GM-CSF receptor signal transduction"— Presentation transcript:

1 Roles of JAK kinases in human GM-CSF receptor signal transduction
Sumiko Watanabe, PhD, Tohru Itoh, MSc, Ken-ichi Arai, MD, PhD  Journal of Allergy and Clinical Immunology  Volume 98, Issue 6, Pages S183-S191 (December 1996) DOI: /S (96) Copyright © 1996 Mosby, Inc. Terms and Conditions

2 FIG. 1 Analysis of requirements of tyrosine residues of hGMR on various signals. A, Schematic drawing of various hGMRβ mutants. B, Activation of c-fos promoter through various C-terminal deletion mutants of hGMRβ. Journal of Allergy and Clinical Immunology  , S183-S191DOI: ( /S (96) ) Copyright © 1996 Mosby, Inc. Terms and Conditions

3 FIG. 1 Analysis of requirements of tyrosine residues of hGMR on various signals. A, Schematic drawing of various hGMRβ mutants. B, Activation of c-fos promoter through various C-terminal deletion mutants of hGMRβ. Journal of Allergy and Clinical Immunology  , S183-S191DOI: ( /S (96) ) Copyright © 1996 Mosby, Inc. Terms and Conditions

4 FIG. 2 Effects of tyrosine residue substitution in hGMRβ on Shc and SHP-2(PTP 1D) phosphorylation. A, c-fos-luciferase activity in the BA/F3 cells expressing hGMRα and various mutants of hGMRβ. BA/F3 cells expressing hGMRα and various mutants of hGMRβ were depleted of mIL-5 for 5 hours and restimulated with hGM-CSF for 5 minutes. Cells were lysed and total proteins (B) or proteins immunoprecipitated by either anti-Shc or anti-SHP-2(PTP 1D) antibodies (C) were analyzed using Western blotting. Journal of Allergy and Clinical Immunology  , S183-S191DOI: ( /S (96) ) Copyright © 1996 Mosby, Inc. Terms and Conditions

5 FIG. 2 Effects of tyrosine residue substitution in hGMRβ on Shc and SHP-2(PTP 1D) phosphorylation. A, c-fos-luciferase activity in the BA/F3 cells expressing hGMRα and various mutants of hGMRβ. BA/F3 cells expressing hGMRα and various mutants of hGMRβ were depleted of mIL-5 for 5 hours and restimulated with hGM-CSF for 5 minutes. Cells were lysed and total proteins (B) or proteins immunoprecipitated by either anti-Shc or anti-SHP-2(PTP 1D) antibodies (C) were analyzed using Western blotting. Journal of Allergy and Clinical Immunology  , S183-S191DOI: ( /S (96) ) Copyright © 1996 Mosby, Inc. Terms and Conditions

6 FIG. 2 Effects of tyrosine residue substitution in hGMRβ on Shc and SHP-2(PTP 1D) phosphorylation. A, c-fos-luciferase activity in the BA/F3 cells expressing hGMRα and various mutants of hGMRβ. BA/F3 cells expressing hGMRα and various mutants of hGMRβ were depleted of mIL-5 for 5 hours and restimulated with hGM-CSF for 5 minutes. Cells were lysed and total proteins (B) or proteins immunoprecipitated by either anti-Shc or anti-SHP-2(PTP 1D) antibodies (C) were analyzed using Western blotting. Journal of Allergy and Clinical Immunology  , S183-S191DOI: ( /S (96) ) Copyright © 1996 Mosby, Inc. Terms and Conditions

7 FIG. 3 Requirements of box1, box2 region for various hGM-CSF activities. A, Effects of deletion of either box1 or box2 on c-fos (A) or c-myc (B) and egr-1 (C) promoter activations by hGM-CSF. BA/F3 cells expressing wild type hGMRα and hGMRβ mutants lacking either the box1 or box2 were transfected by c-fos-luciferase, c-myc -CAT, or egr-1-CAT plasmids. Activation of these promoters by hGM-CSF was analyzed. Journal of Allergy and Clinical Immunology  , S183-S191DOI: ( /S (96) ) Copyright © 1996 Mosby, Inc. Terms and Conditions

8 FIG. 3 Requirements of box1, box2 region for various hGM-CSF activities. A, Effects of deletion of either box1 or box2 on c-fos (A) or c-myc (B) and egr-1 (C) promoter activations by hGM-CSF. BA/F3 cells expressing wild type hGMRα and hGMRβ mutants lacking either the box1 or box2 were transfected by c-fos-luciferase, c-myc -CAT, or egr-1-CAT plasmids. Activation of these promoters by hGM-CSF was analyzed. Journal of Allergy and Clinical Immunology  , S183-S191DOI: ( /S (96) ) Copyright © 1996 Mosby, Inc. Terms and Conditions

9 FIG. 3 Requirements of box1, box2 region for various hGM-CSF activities. A, Effects of deletion of either box1 or box2 on c-fos (A) or c-myc (B) and egr-1 (C) promoter activations by hGM-CSF. BA/F3 cells expressing wild type hGMRα and hGMRβ mutants lacking either the box1 or box2 were transfected by c-fos-luciferase, c-myc -CAT, or egr-1-CAT plasmids. Activation of these promoters by hGM-CSF was analyzed. Journal of Allergy and Clinical Immunology  , S183-S191DOI: ( /S (96) ) Copyright © 1996 Mosby, Inc. Terms and Conditions

10 FIG. 4 Phosphorylation of hGMRβ and cellular proteins through hGMRβ mutants. Total proteins (b) or immunoprecipitants of hGMRβ (a) were electrophoresed and tyrosine phosphorylation was analyzed using Western blotting using anti-phospho-tyrosine antibody or anti β-chain antibody. Journal of Allergy and Clinical Immunology  , S183-S191DOI: ( /S (96) ) Copyright © 1996 Mosby, Inc. Terms and Conditions

11 FIG. 5 Effect of dominant negative JAK2 in various activities by hGMR. A, Phosphorylation of JAK2 and STAT5 in the presence or absence of ΔJAK2. Plasmids encoding hGMRα and β were transfected to BA/F3 cells with either vector control or ΔJAK2. JAK2 or STAT5 proteins were immunoprecipitated and analyzed with Western blotting using anti-phospho tyrosine antibody 4G10. B, Effects of ΔJAK2 for c-myc promoter activation and proliferation. ΔJAK2 was cotransfected with c-myc CAT plasmid in BA/FGMR cells and CAT activity induced by either mIL-3 Shaded Box or hGM-CSF Solid Square was analyzed using diffusion analysis. Plasmids encoding hGMRα and β subunits were transfected to BA/F3 cells with or without ΔJAK2 plasmids and proliferation activity was analyzed using MTT analysis. C, hGM-CSF induced c-fos and egr-1 promoter activation in the presence of dominant negative JAK2. c-fos-luciferase or egr-1-CAT plasmids were transfected to BA/FGMR cells with and without ΔJAK2, and luciferase or CAT activities induced by mIL-3 Shaded Box or hGM-CSF Solid Square were analyzed. Journal of Allergy and Clinical Immunology  , S183-S191DOI: ( /S (96) ) Copyright © 1996 Mosby, Inc. Terms and Conditions

12 FIG. 5 Effect of dominant negative JAK2 in various activities by hGMR. A, Phosphorylation of JAK2 and STAT5 in the presence or absence of ΔJAK2. Plasmids encoding hGMRα and β were transfected to BA/F3 cells with either vector control or ΔJAK2. JAK2 or STAT5 proteins were immunoprecipitated and analyzed with Western blotting using anti-phospho tyrosine antibody 4G10. B, Effects of ΔJAK2 for c-myc promoter activation and proliferation. ΔJAK2 was cotransfected with c-myc CAT plasmid in BA/FGMR cells and CAT activity induced by either mIL-3 Shaded Box or hGM-CSF Solid Square was analyzed using diffusion analysis. Plasmids encoding hGMRα and β subunits were transfected to BA/F3 cells with or without ΔJAK2 plasmids and proliferation activity was analyzed using MTT analysis. C, hGM-CSF induced c-fos and egr-1 promoter activation in the presence of dominant negative JAK2. c-fos-luciferase or egr-1-CAT plasmids were transfected to BA/FGMR cells with and without ΔJAK2, and luciferase or CAT activities induced by mIL-3 Shaded Box or hGM-CSF Solid Square were analyzed. Journal of Allergy and Clinical Immunology  , S183-S191DOI: ( /S (96) ) Copyright © 1996 Mosby, Inc. Terms and Conditions

13 FIG. 5 Effect of dominant negative JAK2 in various activities by hGMR. A, Phosphorylation of JAK2 and STAT5 in the presence or absence of ΔJAK2. Plasmids encoding hGMRα and β were transfected to BA/F3 cells with either vector control or ΔJAK2. JAK2 or STAT5 proteins were immunoprecipitated and analyzed with Western blotting using anti-phospho tyrosine antibody 4G10. B, Effects of ΔJAK2 for c-myc promoter activation and proliferation. ΔJAK2 was cotransfected with c-myc CAT plasmid in BA/FGMR cells and CAT activity induced by either mIL-3 Shaded Box or hGM-CSF Solid Square was analyzed using diffusion analysis. Plasmids encoding hGMRα and β subunits were transfected to BA/F3 cells with or without ΔJAK2 plasmids and proliferation activity was analyzed using MTT analysis. C, hGM-CSF induced c-fos and egr-1 promoter activation in the presence of dominant negative JAK2. c-fos-luciferase or egr-1-CAT plasmids were transfected to BA/FGMR cells with and without ΔJAK2, and luciferase or CAT activities induced by mIL-3 Shaded Box or hGM-CSF Solid Square were analyzed. Journal of Allergy and Clinical Immunology  , S183-S191DOI: ( /S (96) ) Copyright © 1996 Mosby, Inc. Terms and Conditions

14 FIG. 6 Phosphorylation of various mutants and wild type of hGMRβ subunits in COS7 cells in the presence of JAK2. Plasmids encoding wild type or various mutants of hGMRβ were transfected to COS7 cells with JAK2 plasmid and phosphorylation of these receptors was analyzed using immunoprecipitation followed by immunoblotting. Journal of Allergy and Clinical Immunology  , S183-S191DOI: ( /S (96) ) Copyright © 1996 Mosby, Inc. Terms and Conditions

Download ppt "Roles of JAK kinases in human GM-CSF receptor signal transduction"

Similar presentations

Proinflammatory cytokine–induced and chemical mediator–induced IL-8 expression in human bronchial epithelial cells through p38 mitogen-activated protein.

Proinflammatory cytokine–induced and chemical mediator–induced IL-8 expression in human bronchial epithelial cells through p38 mitogen-activated protein.
Volume 56, Issue 5, Pages (November 1999)

Volume 56, Issue 5, Pages (November 1999)
Shin-Ichi Nishikawa, MD, PhD 

Shin-Ichi Nishikawa, MD, PhD 
A novel SHP-1/Grb2–dependent mechanism of negative regulation of cytokine-receptor signaling: contribution of SHP-1 C-terminal tyrosines in cytokine signaling.

A novel SHP-1/Grb2–dependent mechanism of negative regulation of cytokine-receptor signaling: contribution of SHP-1 C-terminal tyrosines in cytokine signaling.
Implications of somatic mutations in the AML1 gene in radiation-associated and therapy-related myelodysplastic syndrome/acute myeloid leukemia by Hironori.

Implications of somatic mutations in the AML1 gene in radiation-associated and therapy-related myelodysplastic syndrome/acute myeloid leukemia by Hironori.
Volume 9, Issue 5, Pages (November 1998)

Volume 9, Issue 5, Pages (November 1998)
Vanessa L. Ott, PhD, Dana C. Fong, PhD, John C. Cambier, PhD 

Vanessa L. Ott, PhD, Dana C. Fong, PhD, John C. Cambier, PhD 
Physical association of Fc receptor γ chain homodimer with IgA receptor  Kan Saito, DMDa, b, Katsuhiro Suzuki, MSa, Hironori Matsuda, BSa, Ko Okumura,

Physical association of Fc receptor γ chain homodimer with IgA receptor  Kan Saito, DMDa, b, Katsuhiro Suzuki, MSa, Hironori Matsuda, BSa, Ko Okumura,
Laurent L'homme, PhD, David Dombrowicz, PhD 

Laurent L'homme, PhD, David Dombrowicz, PhD 
Christopher L. Kepley, PhD, Bridget S. Wilson, PhD, Janet M

Christopher L. Kepley, PhD, Bridget S. Wilson, PhD, Janet M
Roles of the cytoplasmic domains of the α and β subunits of human granulocyte- macrophage colony-stimulating factor receptor  Akihiko Muto, PhDa, Sumiko.

Roles of the cytoplasmic domains of the α and β subunits of human granulocyte- macrophage colony-stimulating factor receptor  Akihiko Muto, PhDa, Sumiko.
The N‐terminus and SH2 domain of SOCS‐1 contribute to inhibition of JAK1 and JAK2 kinase activity. The N‐terminus and SH2 domain of SOCS‐1 contribute to.

The N‐terminus and SH2 domain of SOCS‐1 contribute to inhibition of JAK1 and JAK2 kinase activity. The N‐terminus and SH2 domain of SOCS‐1 contribute to.
Regulation of GM-CSF expression by the transcription factor c-Maf

Regulation of GM-CSF expression by the transcription factor c-Maf
Factors affecting the cytokine production of human T cells stimulated by different modes of activation  Yoshio Harada, MDa, Sumiko Watanabe, PhDa, Hans.

Factors affecting the cytokine production of human T cells stimulated by different modes of activation  Yoshio Harada, MDa, Sumiko Watanabe, PhDa, Hans.
by Fan Dong, and Andrew C. Larner

by Fan Dong, and Andrew C. Larner
Takashi Tanaka, Michelle A. Soriano, Michael J. Grusby  Immunity 

Takashi Tanaka, Michelle A. Soriano, Michael J. Grusby  Immunity 
Lyn Physically Associates With the Erythropoietin Receptor and May Play a Role in Activation of the Stat5 Pathway by Hiroshi Chin, Ayako Arai, Hiroshi.

Lyn Physically Associates With the Erythropoietin Receptor and May Play a Role in Activation of the Stat5 Pathway by Hiroshi Chin, Ayako Arai, Hiroshi.
Signal transduction pathways triggered by the FcϵRIIb receptor (CD23) in human monocytes lead to nuclear factor-κB activation  Rosa M. Ten, MD, PhDa,

Signal transduction pathways triggered by the FcϵRIIb receptor (CD23) in human monocytes lead to nuclear factor-κB activation  Rosa M. Ten, MD, PhDa,
Differential influence of tyrosine residues of the common receptor β subunit on multiple signals induced by human GM-CSF  Tohru Itoh, PhD, Rui Liu, MSc,

Differential influence of tyrosine residues of the common receptor β subunit on multiple signals induced by human GM-CSF  Tohru Itoh, PhD, Rui Liu, MSc,
How cells die: Apoptosis pathways

How cells die: Apoptosis pathways

Similar presentations

Ads by Google