Disabled-2 is a negative regulator of integrin αIIbβ 3-mediated fibrinogen adhesion and cell signaling

Chien Ling Huang, Ju Chien Cheng, Chang Hui Liao, Arnold Stern, Jer Tsong Hsieh, Chi Huei Wang, Hsueh Ling Hsu, Ching Ping Tseng

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45 Scopus citations

Abstract

Disabled-2 (DAB2) is an adapter protein that is up-regulated during megakaryocytic differentiation of hematopoietic cells and is abundantly expressed in platelets. In this study, the role of DAB2 in integrin αIIbβ3-mediated matrix protein fibrinogen adhesion and cell signaling was investigated. In K562 cells differentiating to the megakaryocytic lineage, down-regulation of DAB2 by DAB2 small interfering RNA augmented integrin αIIbβ3 activation and resulted in an increase in cell adhesion to fibrinogen. Ectopic expression of DAB2 reversed the DAB2 small interfering RNA effect or, by itself, decreased fibrinogen adhesion of K562 cells. Mutational analysis revealed that a DAB2 Ser24 phosphorylation mutant (S24A) abrogated the inhibitory function of DAB2. The spatial and temporal association/interaction of DAB2 and platelet integrin αIIbβ3 (CD61) in both megakaryocytic cells and platelets led us to examine the effect of Ser24 phosphorylation on the interaction between DAB2 and integrin β3. Through cellular localization and co-immunoprecipitation analysis, we demonstrate for the first time that Ser24 phosphorylation promotes membrane translocation of DAB2 and its subsequent interaction with integrin β3, thereby defining a mechanism for DAB2 in regulating integrin αIIbβ3 activation and inside-out signaling. Consistent with the effect on fibrinogen adhesion, Ser24 phosphorylation of DAB2 was also involved in the negative regulation of αIIbβ3-induced T cell factor transcriptional activity. In contrast, the S24A mutant acted like wild-type DAB2 and inhibited both β-catenin- and plakoglobin-mediated T cell factor transactivation. Hence, DAB2 elicits distinct regulatory mechanisms in α IIbβ3 and β-catenin/plakoglobin signaling in a Ser24 phosphorylation-dependent and -independent manner, respectively. These findings indicate Ser24 phosphorylation as a molecular basis for DAB2 acting as a negative regulator in α IIbβ3 inside-out signaling and contribute to our understanding of DAB2 in megakaryocytic differentiation and platelet function.

Original languageEnglish (US)
Pages (from-to)42279-42289
Number of pages11
JournalJournal of Biological Chemistry
Volume279
Issue number40
DOIs
StatePublished - Oct 1 2004

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ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Huang, C. L., Cheng, J. C., Liao, C. H., Stern, A., Hsieh, J. T., Wang, C. H., Hsu, H. L., & Tseng, C. P. (2004). Disabled-2 is a negative regulator of integrin αIIbβ 3-mediated fibrinogen adhesion and cell signaling. Journal of Biological Chemistry, 279(40), 42279-42289. https://doi.org/10.1074/jbc.M402540200