The N-terminal SH2 domain of Syk is required for (hem)ITAM, but not integrin, signaling in mouse platelets

Research output: Contribution to journalArticle

Colleges, School and Institutes

Abstract

We have used a novel knockin mouse to investigate the effect of disruption of phosphotyrosine binding of the N-terminal SH2 domain of Syk on platelet activation by GPVI, CLEC-2, and integrin αIIbβ3. The Syk(R41Afl/fl) mouse was crossed to a PF4-Cre(+) mouse to induce expression of the Syk mutant in the megakaryocyte/platelet lineage. Syk(R41Afl/fl;PF4-Cre) mice are born at approximately 50% of the expected frequency and have a similar phenotype to Syk(fl/fl;PF4-Cre) mice, including blood-lymphatic mixing and chyloascites. Anastomosis of the venous and lymphatic vasculatures can be seen in the mesenteric circulation accounting for rapid and continuous mixing of the 2 vasculatures. Platelet activation by CLEC-2 and GPVI is abolished in Syk(R41Afl/fl;PF4-Cre) platelets. Syk phosphorylation on Tyr519/20 is blocked in CLEC-2-stimulated platelets, suggesting a model in which binding of Syk via its N-terminal SH2 domain regulates autophosphorylation. In contrast, outside-in signaling by integrin αIIbβ3 is not altered, but it is inhibited in the presence of inhibitors of Src and Syk tyrosine kinases. These results demonstrate that αIIbβ3 regulates Syk through an ITAM-independent pathway in mice and provide novel insight into the course of events underlying Syk activation and hemITAM phosphorylation by CLEC-2.

Bibliographic note

© 2015 by The American Society of Hematology.

Details

Original languageEnglish
Pages (from-to)144-154
Number of pages11
JournalBlood
Volume125
Issue number1
Early online date28 Oct 2014
Publication statusPublished - 1 Jan 2015

Keywords

  • Amino Acid Motifs, Amino Acid Sequence, Animals, Blood Platelets, Intracellular Signaling Peptides and Proteins, Lectins, C-Type, Mice, Mice, Transgenic, Phosphoproteins, Phosphorylation, Phosphotyrosine, Platelet Activation, Platelet Aggregation, Platelet Glycoprotein GPIIb-IIIa Complex, Platelet Membrane Glycoproteins, Protein Structure, Tertiary, Protein-Tyrosine Kinases, Purpura, Thrombocytopenic, Idiopathic, Signal Transduction, src Homology Domains