Role of vav1- and src-related Tyrosine Kinases in Macrophage Activation by CpG DNA

Stephanie H. Stovall, Ae-Kyung Yi, Elizabeth A. Meals, Ajay Talati, Sandip A. Godambe, B. Keith English

Research output: Contribution to journalArticle

37 Citations (Scopus)

Abstract

Macrophage activation by CpG DNA requires toll-like receptor 9 and the adaptor protein MyD88. Gram-negative bacterial lipopolysaccharide also activates macrophages via a toll-like receptor pathway (TLR-4), but we and others have reported that lipopolysaccharide also stimulates tyrosine phosphorylation in macrophages. Herein we report that exposure of RAW 264.7 murine macrophages to CpG DNA (but not non-CpG DNA) provoked the rapid tyrosine phosphorylation of vav1. PP1, a selective inhibitor of src-related tyrosine kinases, blocked both the CpG DNA-mediated tyrosine phosphorylation of vav1 and the CpG DNA-mediated up-regulation of macrophage tumor necrosis factor secretion and inducible nitric-oxide synthase protein accumulation. Furthermore, we found that the inducible expression of any of three dominant interfering mutants of vav1 (a truncated protein, vavC; a form containing a point mutation in the regulatory tyrosine residue, vavYF174; and a form with an in-frame deletion of six amino acids required for the guanidine nucleotide exchange factor (GEF) activity of vav1 for rac family GTPases, vavGEFmt) consistently inhibited CpG DNA-mediated up-regulation of tumor necrosis factor secretion and inducible nitric-oxide synthase protein accumulation in RAW-TT10 macrophages. Finally, we determined that CpG DNA-mediated up-regulation of NF-κB activity (but not mitogen-activated protein kinase activation) was inhibited by preincubation with PP1 or by expression of the truncated vavC mutant. Taken together, our results indicate that the tyrosine phosphorylation of vav1 by a src-related tyrosine kinase or kinases plays an important role in the macrophage response to CpG DNA.

Original languageEnglish (US)
Pages (from-to)13809-13816
Number of pages8
JournalJournal of Biological Chemistry
Volume279
Issue number14
DOIs
StatePublished - Apr 2 2004

Fingerprint

Macrophage Activation
src-Family Kinases
Macrophages
Protein-Tyrosine Kinases
Chemical activation
Phosphorylation
DNA
Tyrosine
Up-Regulation
Nitric Oxide Synthase Type II
Lipopolysaccharides
Tumor Necrosis Factor-alpha
Myeloid Differentiation Factor 88
Toll-Like Receptor 9
Proteins
Toll-Like Receptors
GTP Phosphohydrolases
Guanidine
Mitogen-Activated Protein Kinases
Point Mutation

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Role of vav1- and src-related Tyrosine Kinases in Macrophage Activation by CpG DNA. / Stovall, Stephanie H.; Yi, Ae-Kyung; Meals, Elizabeth A.; Talati, Ajay; Godambe, Sandip A.; English, B. Keith.

In: Journal of Biological Chemistry, Vol. 279, No. 14, 02.04.2004, p. 13809-13816.

Research output: Contribution to journalArticle

Stovall, Stephanie H. ; Yi, Ae-Kyung ; Meals, Elizabeth A. ; Talati, Ajay ; Godambe, Sandip A. ; English, B. Keith. / Role of vav1- and src-related Tyrosine Kinases in Macrophage Activation by CpG DNA. In: Journal of Biological Chemistry. 2004 ; Vol. 279, No. 14. pp. 13809-13816.
@article{fc077930eb034c978190e7a43feb32ec,
title = "Role of vav1- and src-related Tyrosine Kinases in Macrophage Activation by CpG DNA",
abstract = "Macrophage activation by CpG DNA requires toll-like receptor 9 and the adaptor protein MyD88. Gram-negative bacterial lipopolysaccharide also activates macrophages via a toll-like receptor pathway (TLR-4), but we and others have reported that lipopolysaccharide also stimulates tyrosine phosphorylation in macrophages. Herein we report that exposure of RAW 264.7 murine macrophages to CpG DNA (but not non-CpG DNA) provoked the rapid tyrosine phosphorylation of vav1. PP1, a selective inhibitor of src-related tyrosine kinases, blocked both the CpG DNA-mediated tyrosine phosphorylation of vav1 and the CpG DNA-mediated up-regulation of macrophage tumor necrosis factor secretion and inducible nitric-oxide synthase protein accumulation. Furthermore, we found that the inducible expression of any of three dominant interfering mutants of vav1 (a truncated protein, vavC; a form containing a point mutation in the regulatory tyrosine residue, vavYF174; and a form with an in-frame deletion of six amino acids required for the guanidine nucleotide exchange factor (GEF) activity of vav1 for rac family GTPases, vavGEFmt) consistently inhibited CpG DNA-mediated up-regulation of tumor necrosis factor secretion and inducible nitric-oxide synthase protein accumulation in RAW-TT10 macrophages. Finally, we determined that CpG DNA-mediated up-regulation of NF-κB activity (but not mitogen-activated protein kinase activation) was inhibited by preincubation with PP1 or by expression of the truncated vavC mutant. Taken together, our results indicate that the tyrosine phosphorylation of vav1 by a src-related tyrosine kinase or kinases plays an important role in the macrophage response to CpG DNA.",
author = "Stovall, {Stephanie H.} and Ae-Kyung Yi and Meals, {Elizabeth A.} and Ajay Talati and Godambe, {Sandip A.} and English, {B. Keith}",
year = "2004",
month = "4",
day = "2",
doi = "10.1074/jbc.M311434200",
language = "English (US)",
volume = "279",
pages = "13809--13816",
journal = "Journal of Biological Chemistry",
issn = "0021-9258",
publisher = "American Society for Biochemistry and Molecular Biology Inc.",
number = "14",

}

TY - JOUR

T1 - Role of vav1- and src-related Tyrosine Kinases in Macrophage Activation by CpG DNA

AU - Stovall, Stephanie H.

AU - Yi, Ae-Kyung

AU - Meals, Elizabeth A.

AU - Talati, Ajay

AU - Godambe, Sandip A.

AU - English, B. Keith

PY - 2004/4/2

Y1 - 2004/4/2

N2 - Macrophage activation by CpG DNA requires toll-like receptor 9 and the adaptor protein MyD88. Gram-negative bacterial lipopolysaccharide also activates macrophages via a toll-like receptor pathway (TLR-4), but we and others have reported that lipopolysaccharide also stimulates tyrosine phosphorylation in macrophages. Herein we report that exposure of RAW 264.7 murine macrophages to CpG DNA (but not non-CpG DNA) provoked the rapid tyrosine phosphorylation of vav1. PP1, a selective inhibitor of src-related tyrosine kinases, blocked both the CpG DNA-mediated tyrosine phosphorylation of vav1 and the CpG DNA-mediated up-regulation of macrophage tumor necrosis factor secretion and inducible nitric-oxide synthase protein accumulation. Furthermore, we found that the inducible expression of any of three dominant interfering mutants of vav1 (a truncated protein, vavC; a form containing a point mutation in the regulatory tyrosine residue, vavYF174; and a form with an in-frame deletion of six amino acids required for the guanidine nucleotide exchange factor (GEF) activity of vav1 for rac family GTPases, vavGEFmt) consistently inhibited CpG DNA-mediated up-regulation of tumor necrosis factor secretion and inducible nitric-oxide synthase protein accumulation in RAW-TT10 macrophages. Finally, we determined that CpG DNA-mediated up-regulation of NF-κB activity (but not mitogen-activated protein kinase activation) was inhibited by preincubation with PP1 or by expression of the truncated vavC mutant. Taken together, our results indicate that the tyrosine phosphorylation of vav1 by a src-related tyrosine kinase or kinases plays an important role in the macrophage response to CpG DNA.

AB - Macrophage activation by CpG DNA requires toll-like receptor 9 and the adaptor protein MyD88. Gram-negative bacterial lipopolysaccharide also activates macrophages via a toll-like receptor pathway (TLR-4), but we and others have reported that lipopolysaccharide also stimulates tyrosine phosphorylation in macrophages. Herein we report that exposure of RAW 264.7 murine macrophages to CpG DNA (but not non-CpG DNA) provoked the rapid tyrosine phosphorylation of vav1. PP1, a selective inhibitor of src-related tyrosine kinases, blocked both the CpG DNA-mediated tyrosine phosphorylation of vav1 and the CpG DNA-mediated up-regulation of macrophage tumor necrosis factor secretion and inducible nitric-oxide synthase protein accumulation. Furthermore, we found that the inducible expression of any of three dominant interfering mutants of vav1 (a truncated protein, vavC; a form containing a point mutation in the regulatory tyrosine residue, vavYF174; and a form with an in-frame deletion of six amino acids required for the guanidine nucleotide exchange factor (GEF) activity of vav1 for rac family GTPases, vavGEFmt) consistently inhibited CpG DNA-mediated up-regulation of tumor necrosis factor secretion and inducible nitric-oxide synthase protein accumulation in RAW-TT10 macrophages. Finally, we determined that CpG DNA-mediated up-regulation of NF-κB activity (but not mitogen-activated protein kinase activation) was inhibited by preincubation with PP1 or by expression of the truncated vavC mutant. Taken together, our results indicate that the tyrosine phosphorylation of vav1 by a src-related tyrosine kinase or kinases plays an important role in the macrophage response to CpG DNA.

UR - http://www.scopus.com/inward/record.url?scp=1842791354&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=1842791354&partnerID=8YFLogxK

U2 - 10.1074/jbc.M311434200

DO - 10.1074/jbc.M311434200

M3 - Article

VL - 279

SP - 13809

EP - 13816

JO - Journal of Biological Chemistry

JF - Journal of Biological Chemistry

SN - 0021-9258

IS - 14

ER -