A role for Grb2-associated binder-1 in growth hormone signaling

Sung Oh Kim, Kimberly Loesch, Xiangdong Wang, Jing Jiang, Lin Mei, Jess M. Cunnick, Jie Wu, Stuart J. Frank

Research output: Contribution to journalArticle

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Abstract

GH signaling begins with activation of the GH receptor (GHR)-associated cytoplasmic tyrosine kinase, Janus kinase-2. GH-induced Janus kinase-2 activation leads to engagement of several signaling pathways, including the extracellular signal-regulated kinase (ERK), mitogen-activated protein kinase, phosphoinositol 3-kinase, and signal transducer and activator of transcription-5 (STAT5) pathways. Previous work suggests that ERK activation in response to GH may be modulated by several proteins acting as docking molecules, including the epidermal growth factor receptor (EGFR) and insulin receptor substrate-1. In this study we investigate potential roles for the pleckstrin homology (PH) domain-containing insulin receptor substrate-like protein, Grb-2-associated binder-1 (Gab1), in GH signaling. We find in 3T3-F442A preadipocytes that GH promotes tyrosine phosphorylation of Gab1 and its association with SHP2, an Src homology 2-containing cytoplasmic tyrosine phosphatase. The Grb2 adapter protein, in contrast, is specifically coimmunoprecipitated with Gab1, even in the absence of GH exposure. Using a COS-7 cell transient reconstitution system, we observed that GH-induced Gab1 tyrosine phosphorylation is dependent on the Gab1 PH domain, whereas GH-induced coimmunoprecipitation of SHP2 requires tyrosine 627 of Gab1, as previously reported for EGF-induced Gab1-SHP2 association. Deletion of the Gab1 PH domain significantly attenuates GH-induced ERK activation and trans-activation of a c-fos enhancer-driven reporter construct compared with wild-type Gab1 in this system. In contrast, GH-induced STAT5 tyrosine phosphorylation and STAT5-dependent trans-activation are similar in cells expressing wild-type or PH domain-deleted Gab1. Notably, neither the ERK nor the STAT5 GH-dependent signaling outcome is affected by expression of the Gab1 mutant with tyrosine 627 changed to phenylalanine. Finally, we observed GH-dependent translocation of a wild-type, but not a PH domain-deleted, Gab1-green fluorescent protein chimera from the cytoplasm to the plasma membrane. Our results suggest selective involvement of Gab1 in GH-induced ERK activation and implicate the Gab1 PH domain as critical in this involvement.

Original languageEnglish (US)
Pages (from-to)4856-4867
Number of pages12
JournalEndocrinology
Volume143
Issue number12
DOIs
StatePublished - Dec 1 2002

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STAT5 Transcription Factor
Extracellular Signal-Regulated MAP Kinases
Growth Hormone
Tyrosine
Janus Kinase 2
Insulin Receptor Substrate Proteins
Phosphorylation
MAP Kinase Kinase Kinase 3
COS Cells
Cytoplasmic and Nuclear Receptors
Green Fluorescent Proteins
Phenylalanine
Phosphoric Monoester Hydrolases
Epidermal Growth Factor Receptor
Epidermal Growth Factor
Protein-Tyrosine Kinases
Pleckstrin Homology Domains
Cytoplasm
Proteins
Cell Membrane

ASJC Scopus subject areas

  • Endocrinology

Cite this

Kim, S. O., Loesch, K., Wang, X., Jiang, J., Mei, L., Cunnick, J. M., ... Frank, S. J. (2002). A role for Grb2-associated binder-1 in growth hormone signaling. Endocrinology, 143(12), 4856-4867. https://doi.org/10.1210/en.2002-220565

A role for Grb2-associated binder-1 in growth hormone signaling. / Kim, Sung Oh; Loesch, Kimberly; Wang, Xiangdong; Jiang, Jing; Mei, Lin; Cunnick, Jess M.; Wu, Jie; Frank, Stuart J.

In: Endocrinology, Vol. 143, No. 12, 01.12.2002, p. 4856-4867.

Research output: Contribution to journalArticle

Kim, SO, Loesch, K, Wang, X, Jiang, J, Mei, L, Cunnick, JM, Wu, J & Frank, SJ 2002, 'A role for Grb2-associated binder-1 in growth hormone signaling', Endocrinology, vol. 143, no. 12, pp. 4856-4867. https://doi.org/10.1210/en.2002-220565
Kim SO, Loesch K, Wang X, Jiang J, Mei L, Cunnick JM et al. A role for Grb2-associated binder-1 in growth hormone signaling. Endocrinology. 2002 Dec 1;143(12):4856-4867. https://doi.org/10.1210/en.2002-220565
Kim, Sung Oh ; Loesch, Kimberly ; Wang, Xiangdong ; Jiang, Jing ; Mei, Lin ; Cunnick, Jess M. ; Wu, Jie ; Frank, Stuart J. / A role for Grb2-associated binder-1 in growth hormone signaling. In: Endocrinology. 2002 ; Vol. 143, No. 12. pp. 4856-4867.
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