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First published online 28 March 2006
doi: 10.1242/jcs.02845

Journal of Cell Science 119, 1666-1676 (2006)
Published by The Company of Biologists 2006
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Research Article

Interaction of SH2-Bß with RET is involved in signaling of GDNF-induced neurite outgrowth

Yong Zhang*, Wei Zhu*, Yong-Gang Wang, Xiu-Jie Liu, Li Jiao, Xuan Liu, Zhao-Huan Zhang, Chang-Lin Lu and Cheng He{ddagger}

Department of Neurobiology, Second Military Medical University, Shanghai, 200433, P. R. of China

{ddagger} Author for correspondence (e-mail: chenghe{at}online.sh.cn)

Accepted 20 December 2005

RET receptor signalling is essential for glial-cell-line-derived neurotrophic factor (GDNF)-induced survival and differentiation of various neurons such as mesencephalic neurons. To identify proteins that mediate RET-dependent signaling, yeast two-hybrid screening was performed with the intracellular domain of RET as bait. We identified a new interaction between RET and the adapter protein SH2-Bß. Upon GDNF stimulation of PC12-GFR{alpha}1-RET cells (that stably overexpress GDNF receptor {alpha}1 and RET), wild-type SH2-Bß co-immunoprecipitated with RET, whereas the dominant-negative SH2-Bß mutant R555E did not. RET interacted with endogenous SH2-Bß both in PC12-GFR{alpha}1-RET cells and in rat tissues. Mutagenesis analysis revealed that Tyr981 within the intracellular domain of RET was crucial for the interaction with SH2-Bß. Morphological evidence showed that SH2-Bß and RET colocalized in mesencephalic neurons. Furthermore, functional analysis indicated that overexpression of SH2-Bß facilitated GDNF-induced neurite outgrowth in both PC12-GFR{alpha}1-RET cells and cultured mesencephalic neurons, whereas the mutant R555E inhibited the effect. Moreover, inhibition of SH2-Bß expression by RNA interference caused a significant decrease of GDNF-induced neuronal differentiation in PC12-GFR{alpha}1-RET cells. Taken together, our results suggest that SH2-Bß is a new signaling molecule involved in GDNF-induced neurite outgrowth.

Key words: RET, GDNF, SH2-Bß, Interaction, Neurite outgrowth


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© The Company of Biologists Ltd 2006