The pleckstrin homology domain of Gab-2 is required for optimal interleukin-3 signalsome-mediated responses

C E Edmead, B C Fox, C Stace, N Ktistakis, M J Welham

Research output: Contribution to journalArticle

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Abstract

The adaptor protein Gab-2 coordinates the assembly of the IL-3 signalsome comprising Gab-2, Grb2, Shc, SHP-2 and PI3K. To investigate the role of the pleckstrin homology domain of Gab-2 in this process, epitope-tagged wild type Gab-2 (WTGab-2), Gab-2 lacking its PH domain (ΔPHGab-2) and the Gab-2 PH domain alone (PHGab-2) were inducibly expressed in IL-3-dependent BaF/3 cells. Expression of ΔPHGab-2 reduced IL-3-dependent proliferation and long-term activation of ERK1 and 2 and PKB by IL-3. While we demonstrate that the Gab-2 PH domain can bind PI(3,4,5)P3, it is dispensable for Gab-2 membrane localisation, tyrosine phosphorylation and signalsome formation. Rather, the proline-rich motifs of Gab-2 appear to contribute to the constitutive membrane localisation we observe, independently of tyrosine phosphorylation or the PH domain. Taken together, these findings suggest that once Gab-2 is tyrosine phosphorylated its PH domain is required for the optimal stabilisation of the signalsome, enabling full activation of downstream signals.
Original languageEnglish
Pages (from-to)1147-1155
Number of pages9
JournalCellular Signalling
Volume18
Issue number8
DOIs
Publication statusPublished - 2006

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Interleukin-3
Tyrosine
Phosphorylation
Membranes
Phosphatidylinositol 3-Kinases
Proline
Pleckstrin Homology Domains
Epitopes
Proteins

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The pleckstrin homology domain of Gab-2 is required for optimal interleukin-3 signalsome-mediated responses. / Edmead, C E; Fox, B C; Stace, C; Ktistakis, N; Welham, M J.

In: Cellular Signalling, Vol. 18, No. 8, 2006, p. 1147-1155.

Research output: Contribution to journalArticle

Edmead, C E ; Fox, B C ; Stace, C ; Ktistakis, N ; Welham, M J. / The pleckstrin homology domain of Gab-2 is required for optimal interleukin-3 signalsome-mediated responses. In: Cellular Signalling. 2006 ; Vol. 18, No. 8. pp. 1147-1155.
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AB - The adaptor protein Gab-2 coordinates the assembly of the IL-3 signalsome comprising Gab-2, Grb2, Shc, SHP-2 and PI3K. To investigate the role of the pleckstrin homology domain of Gab-2 in this process, epitope-tagged wild type Gab-2 (WTGab-2), Gab-2 lacking its PH domain (ΔPHGab-2) and the Gab-2 PH domain alone (PHGab-2) were inducibly expressed in IL-3-dependent BaF/3 cells. Expression of ΔPHGab-2 reduced IL-3-dependent proliferation and long-term activation of ERK1 and 2 and PKB by IL-3. While we demonstrate that the Gab-2 PH domain can bind PI(3,4,5)P3, it is dispensable for Gab-2 membrane localisation, tyrosine phosphorylation and signalsome formation. Rather, the proline-rich motifs of Gab-2 appear to contribute to the constitutive membrane localisation we observe, independently of tyrosine phosphorylation or the PH domain. Taken together, these findings suggest that once Gab-2 is tyrosine phosphorylated its PH domain is required for the optimal stabilisation of the signalsome, enabling full activation of downstream signals.

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