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The Ret receptor protein tyrosine kinase associates with the SH2-containing adapter protein Grb10
A Pandey1, H Duan, P P Di Fiore
1Department of Pathology, University of Michigan Medical School, Ann Arbor 48109-0602, USA.
Abstract:
Ret is a receptor protein tyrosine kinase that has been implicated in the development of the enteric nervous, endocrine, and renal systems. Mutations associated with multiple endocrine neoplasia types 2A and 2B (MEN 2A and 2B) have been shown to activate the intrinsic kinase and transforming ability of ret (Santoro, M., Carlomagno, F., Romano, A., Bottaro, D. P., Dathan, N. A., Grieco, M., Fusco, A., Vecchio, G., Matoskova, B., Kraus, M. H., and Paolo DiFiore, P. (1995) Science 267, 381-383). Using the cytoplasmic domain of Ret as bait in a yeast two-hybrid screen of a mouse embryonic library, it was discovered that the src homology 2 (SH2) domain containing protein Grb10 bound Ret. Grb10 belongs to an emerging family of SH2 containing adapter proteins, the prototypical member being Grb7. Using glutathione S-transferase fusion proteins, it was demonstrated that the SH2 domain of Grb10 specifically interacted with Ret. Additionally, using an EGFR/Ret chimera, it was shown that Grb10 bound Ret in an activation dependent manner in vivo. This is the first description of a receptor protein tyrosine kinase that utilizes Grb10 as a signaling intermediate.
Insights
The Ret receptor tyrosine kinase interacts with the Grb10 adapter protein. This discovery reveals a novel signaling pathway involving Grb10 in Ret-mediated cellular processes.
Area of Science:
- Molecular Biology
- Cell Signaling
- Biochemistry
Background:
- Ret is a receptor protein tyrosine kinase crucial for development.
- Mutations in Ret are linked to Multiple Endocrine Neoplasia (MEN) types 2A and 2B.
- Activated Ret exhibits enhanced kinase and transforming abilities.
Purpose of the Study:
- To identify novel signaling partners of Ret.
- To investigate the interaction between Ret and Grb10.
- To elucidate the role of Grb10 in Ret signaling.
Main Methods:
- Yeast two-hybrid screening using the Ret cytoplasmic domain.
- Glutathione S-transferase (GST) pull-down assays.
- EGFR/Ret chimera experiments to study in vivo binding.
Main Results:
- Grb10, an SH2 domain-containing protein, was identified as a Ret-binding partner.
- The SH2 domain of Grb10 specifically interacts with Ret.
- Grb10 binding to Ret is activation-dependent in vivo.
Conclusions:
- Grb10 acts as a novel signaling intermediate for Ret.
- This identifies a new pathway in Ret receptor tyrosine kinase signaling.
- The findings provide insights into the molecular mechanisms of Ret function.