The Vav binding site of the non-receptor tyrosine kinase Syk at Tyr 348 is critical for β2 integrin (CD11/CD18)-mediated neutrophil migration

Jurgen Schymeinsky, Anca Sindrilaru, David Frommhold, Markus Sperandio, Ronald Gerstl, Cornelia Then, A. Mócsai, Karin Scharffetter-Kochanek, Barbara Walzog

Research output: Contribution to journalArticle

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Abstract

Leukocyte adhesion via β2 integrins (CD11/CD18) activates the tyrosine kinase Syk. We found that Syk was enriched at the lamellipodium during N-formyl-Met-Leu-Phe-induced migration of neutrophil-like differentiated HL-60 cells. Here, Syk colocalized with Vav, a guanine nucleotide exchange factor for Rac and Cdc42. The enrichment of Syk at the lamellipodium and its colocalization with Vav were absent upon expression of a Syk kinase-dead mutant (Syk K402R) or a Syk mutant lacking the binding site of Vav (Syk Y348F). Live cell imaging revealed that both mutations resulted in excessive lamellipodium formation and severely compromised migration compared with control cells. Similar results were obtained upon down-regulation of Syk by RNA interference (RNAi) technique as well as in Syk-/- neutrophils from wild-type mice reconstituted with Syk-/- bone marrow. A pivotal role of Syk in vivo was demonstrated in the Arthus reaction, where neutrophil extravasation, edema formation, and hemorrhage were profoundly diminished in Syk-/- bone marrow chimeras compared with those in control animals. In the inflamed cremaster muscle, Syk-/- neutrophils revealed a defect in adhesion and migration. These findings indicate that Syk is critical for β2 integrin-mediated neutrophil migration in vitro and plays a fundamental role in neutrophil recruitment during the inflammatory response in vivo.

Original languageEnglish
Pages (from-to)3919-3927
Number of pages9
JournalBlood
Volume108
Issue number12
DOIs
Publication statusPublished - Dec 1 2006

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Integrins
Protein-Tyrosine Kinases
Bone
Neutrophils
Adhesion
Binding Sites
Pseudopodia
Guanine Nucleotide Exchange Factors
Muscle
Animals
Phosphotransferases
Proto-Oncogene Proteins c-vav
RNA
Imaging techniques
Defects
Bone Marrow
Arthus Reaction
Abdominal Muscles
Neutrophil Infiltration
HL-60 Cells

ASJC Scopus subject areas

  • Hematology

Cite this

Schymeinsky, J., Sindrilaru, A., Frommhold, D., Sperandio, M., Gerstl, R., Then, C., ... Walzog, B. (2006). The Vav binding site of the non-receptor tyrosine kinase Syk at Tyr 348 is critical for β2 integrin (CD11/CD18)-mediated neutrophil migration. Blood, 108(12), 3919-3927. https://doi.org/10.1182/blood-2005-12-030387

The Vav binding site of the non-receptor tyrosine kinase Syk at Tyr 348 is critical for β2 integrin (CD11/CD18)-mediated neutrophil migration. / Schymeinsky, Jurgen; Sindrilaru, Anca; Frommhold, David; Sperandio, Markus; Gerstl, Ronald; Then, Cornelia; Mócsai, A.; Scharffetter-Kochanek, Karin; Walzog, Barbara.

In: Blood, Vol. 108, No. 12, 01.12.2006, p. 3919-3927.

Research output: Contribution to journalArticle

Schymeinsky, J, Sindrilaru, A, Frommhold, D, Sperandio, M, Gerstl, R, Then, C, Mócsai, A, Scharffetter-Kochanek, K & Walzog, B 2006, 'The Vav binding site of the non-receptor tyrosine kinase Syk at Tyr 348 is critical for β2 integrin (CD11/CD18)-mediated neutrophil migration', Blood, vol. 108, no. 12, pp. 3919-3927. https://doi.org/10.1182/blood-2005-12-030387
Schymeinsky, Jurgen ; Sindrilaru, Anca ; Frommhold, David ; Sperandio, Markus ; Gerstl, Ronald ; Then, Cornelia ; Mócsai, A. ; Scharffetter-Kochanek, Karin ; Walzog, Barbara. / The Vav binding site of the non-receptor tyrosine kinase Syk at Tyr 348 is critical for β2 integrin (CD11/CD18)-mediated neutrophil migration. In: Blood. 2006 ; Vol. 108, No. 12. pp. 3919-3927.
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abstract = "Leukocyte adhesion via β2 integrins (CD11/CD18) activates the tyrosine kinase Syk. We found that Syk was enriched at the lamellipodium during N-formyl-Met-Leu-Phe-induced migration of neutrophil-like differentiated HL-60 cells. Here, Syk colocalized with Vav, a guanine nucleotide exchange factor for Rac and Cdc42. The enrichment of Syk at the lamellipodium and its colocalization with Vav were absent upon expression of a Syk kinase-dead mutant (Syk K402R) or a Syk mutant lacking the binding site of Vav (Syk Y348F). Live cell imaging revealed that both mutations resulted in excessive lamellipodium formation and severely compromised migration compared with control cells. Similar results were obtained upon down-regulation of Syk by RNA interference (RNAi) technique as well as in Syk-/- neutrophils from wild-type mice reconstituted with Syk-/- bone marrow. A pivotal role of Syk in vivo was demonstrated in the Arthus reaction, where neutrophil extravasation, edema formation, and hemorrhage were profoundly diminished in Syk-/- bone marrow chimeras compared with those in control animals. In the inflamed cremaster muscle, Syk-/- neutrophils revealed a defect in adhesion and migration. These findings indicate that Syk is critical for β2 integrin-mediated neutrophil migration in vitro and plays a fundamental role in neutrophil recruitment during the inflammatory response in vivo.",
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AU - Sperandio, Markus

AU - Gerstl, Ronald

AU - Then, Cornelia

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