Transforming growth factor β activates Smad2 in the absence of receptor endocytosis

Zhongxian Lu, James T. Murray, Wenjie Luo, Hongling Li, Xiaoping Wu, Huaxi Xu, Jonathan M. Backer, Ye Guang Chen

Research output: Contribution to journalArticle

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Abstract

Like many other cell surface receptors, transforming growth factor β (TGF-β) receptors are internalized upon ligand stimulation. Given that the signaling-facilitating molecules Smad anchor for receptor activation (SARA) and Hrs are mainly localized in early endosomes, it was unclear whether receptor internalization is required for Smad2 activation. Using reversible biotin labeling, we directly monitored internalization of the TGF-β type I receptor. Our data indicate that TGF-β type I receptor is endocytosed via a clathrin-dependent mechanism and is effectively blocked by depletion of intracellular potassium or by expression of a mutant dynamin (K44A). However, blockage of receptor endocytosis by these two means has no effect on TGF-β-mediated Smad2 activation. Furthermore, TGF-β-induced Smad2 activation was unaffected by inhibition of hVPS34 activity with wortmannin or inhibitory anti-hVPS34 antibodies. Finally, we demonstrated that Smad2 interacted with cell surface receptors and that a SARA binding-deficient Smad2 mutant was phosphorylated by the receptors. Thus, our findings suggest that receptor endocytosis is dispersible for TGF-β-mediated activation of Smad2 and that this activation can be mediated by both SARA-dependent and -independent mechanisms.

Original languageEnglish (US)
Pages (from-to)29363-29368
Number of pages6
JournalJournal of Biological Chemistry
Volume277
Issue number33
DOIs
StatePublished - Aug 16 2002

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Transforming Growth Factors
Endocytosis
Chemical activation
Growth Factor Receptors
Anchors
Cell Surface Receptors
Dynamins
Clathrin
Endosomes
Biotin
Anti-Idiotypic Antibodies
Potassium
Labeling
Ligands
Molecules

ASJC Scopus subject areas

  • Biochemistry

Cite this

Lu, Z., Murray, J. T., Luo, W., Li, H., Wu, X., Xu, H., ... Chen, Y. G. (2002). Transforming growth factor β activates Smad2 in the absence of receptor endocytosis. Journal of Biological Chemistry, 277(33), 29363-29368. https://doi.org/10.1074/jbc.M203495200

Transforming growth factor β activates Smad2 in the absence of receptor endocytosis. / Lu, Zhongxian; Murray, James T.; Luo, Wenjie; Li, Hongling; Wu, Xiaoping; Xu, Huaxi; Backer, Jonathan M.; Chen, Ye Guang.

In: Journal of Biological Chemistry, Vol. 277, No. 33, 16.08.2002, p. 29363-29368.

Research output: Contribution to journalArticle

Lu, Zhongxian ; Murray, James T. ; Luo, Wenjie ; Li, Hongling ; Wu, Xiaoping ; Xu, Huaxi ; Backer, Jonathan M. ; Chen, Ye Guang. / Transforming growth factor β activates Smad2 in the absence of receptor endocytosis. In: Journal of Biological Chemistry. 2002 ; Vol. 277, No. 33. pp. 29363-29368.
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