Regulation of Microtubule Nucleation in Mouse Bone Marrow-Derived Mast Cells by Protein Tyrosine Phosphatase SHP-1

• Published in: Cells (Basel, Switzerland) Vol. 8; no. 4; p. 345
• Main Authors: Klebanovych, Anastasiya, Sládková, Vladimíra, Sulimenko, Tetyana, Vosecká, Věra, Čapek, Martin, Dráberová, Eduarda, Dráber, Pavel, Sulimenko, Vadym
• Format: Journal Article
• Language: English
• Published: Switzerland MDPI AG 11.04.2019; MDPI
• Subjects: Animals; Bone marrow; bone marrow-derived mast cells; Cell activation; Cell Degranulation; Chemokines; CRISPR; Cytokines; Degranulation; HEK293 Cells; Homology; Humans; Immunoprecipitation; Inflammation; Kinases; Laboratories; Mast cells; Mast Cells - cytology; Mast Cells - metabolism; MCF-7 Cells; Mice; microtubule nucleation; Microtubules - metabolism; Molecular modelling; Nucleation; Phosphatase; Phosphorylation; Plasmids; Protein Tyrosine Phosphatase, Non-Receptor Type 6 - antagonists & inhibitors; Protein Tyrosine Phosphatase, Non-Receptor Type 6 - physiology; Protein-tyrosine kinase; Protein-tyrosine-phosphatase; Proteins; SHP-1 protein; SHP-1 tyrosine phosphatase; Syk Kinase - metabolism; Syk protein; Tubulin; Tubulin - metabolism; γ-tubulin complexes; United States > US; Czech Republic; Germany; microtubule nucleation; bone marrow-derived mast cells; γ-tubulin complexes; SHP-1 tyrosine phosphatase; cell activation
• ISSN: 2073-4409; 2073-4409
• DOI: 10.3390/cells8040345

The antigen-mediated activation of mast cells initiates signaling events leading to their degranulation, to the release of inflammatory mediators, and to the synthesis of cytokines and chemokines. Although rapid and transient microtubule reorganization during activation has been described, the molecular mechanisms that control their rearrangement are largely unknown. Microtubule nucleation is mediated by γ-tubulin complexes. In this study, we report on the regulation of microtubule nucleation in bone marrow-derived mast cells (BMMCs) by Src homology 2 (SH2) domain-containing protein tyrosine phosphatase 1 (SHP-1; Ptpn6). Reciprocal immunoprecipitation experiments and pull-down assays revealed that SHP-1 is present in complexes containing γ-tubulin complex proteins and protein tyrosine kinase Syk. Microtubule regrowth experiments in cells with deleted SHP-1 showed a stimulation of microtubule nucleation, and phenotypic rescue experiments confirmed that SHP-1 represents a negative regulator of microtubule nucleation in BMMCs. Moreover, the inhibition of the SHP-1 activity by inhibitors TPI-1 and NSC87877 also augmented microtubule nucleation. The regulation was due to changes in γ-tubulin accumulation. Further experiments with antigen-activated cells showed that the deletion of SHP-1 stimulated the generation of microtubule protrusions, the activity of Syk kinase, and degranulation. Our data suggest a novel mechanism for the suppression of microtubule formation in the later stages of mast cell activation.