Serine phosphorylation differentially affects RhoA binding to effectors

Implications to NGF-induced neurite outgrowth

Nóra Nusser, Elvira Gosmanova, Natalia Makarova, Yuko Fujiwara, Linda Yang, Fukun Guo, Yongneng Luo, Yi Zheng, Gabor Tigyi

Research output: Contribution to journalArticle

39 Citations (Scopus)

Abstract

Activation of RhoA prevents NGF-induced outgrowth and causes retraction of neurites in neuronal cells, including PC12 cells. Despite its inhibitory effect on neurite outgrowth, NGF activates GTP loading of and effector binding to RhoA, setting up an apparent contradiction. According to the molecular switch hypothesis of GTPase function GTP-loading of RhoA should be sufficient to activate its effectors uniformly. However, when monitoring NGF-induced binding of GTP-RhoA to multiple targets, we noted differential interactions with its effectors. We found that NGF elicits a protein kinase A-mediated phosphorylation of RhoA on serine 188 , which renders it unable to bind to Rho-associated kinase (ROK), whereas it retains the ability to interact with other RhoA targets including rhotekin, mDia-1 and PKN. We show in vitro and in vivo that phosphorylation of serine 188 represents an additional switch, capable of directing signals among effector pathways. In the context of PC12 cell differentiation, NGF-induced phosphorylation of RhoA on serine 188 prevents it from interacting with ROK, which would otherwise block neurite outgrowth. Transfection of RhoA S188A mutant into PC12 cells prevents NGF-induced neurite outgrowth, just like constitutively activated RhoA 14V does, indicating the requirement of this phosphorylation site. Replacement of serine 188 with the phosphomimetic glutamate residue in RhoA V14/S188E selectively impairs interaction with ROK and when transfected into PC12 cells restores NGF-induced neurite outgrowth. Therefore, phosphorylation of serine 188 may serve as a novel secondary switch of RhoA capable of overriding GTP-binding-elicited effector activation to a subset of targets such as ROK, which interact with the C-terminus of RhoA.

Original languageEnglish (US)
Pages (from-to)704-714
Number of pages11
JournalCellular Signalling
Volume18
Issue number5
DOIs
StatePublished - May 1 2006

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Nerve Growth Factor
Serine
Phosphorylation
rho-Associated Kinases
PC12 Cells
Guanosine Triphosphate
GTP Phosphohydrolases
Neurites
Cyclic AMP-Dependent Protein Kinases
Neuronal Outgrowth
Transfection
Cell Differentiation
Glutamic Acid

All Science Journal Classification (ASJC) codes

  • Cell Biology

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Serine phosphorylation differentially affects RhoA binding to effectors : Implications to NGF-induced neurite outgrowth. / Nusser, Nóra; Gosmanova, Elvira; Makarova, Natalia; Fujiwara, Yuko; Yang, Linda; Guo, Fukun; Luo, Yongneng; Zheng, Yi; Tigyi, Gabor.

In: Cellular Signalling, Vol. 18, No. 5, 01.05.2006, p. 704-714.

Research output: Contribution to journalArticle

Nusser, Nóra ; Gosmanova, Elvira ; Makarova, Natalia ; Fujiwara, Yuko ; Yang, Linda ; Guo, Fukun ; Luo, Yongneng ; Zheng, Yi ; Tigyi, Gabor. / Serine phosphorylation differentially affects RhoA binding to effectors : Implications to NGF-induced neurite outgrowth. In: Cellular Signalling. 2006 ; Vol. 18, No. 5. pp. 704-714.
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