Phosphorylation and Activation of RhoA by ERK in Response to Epidermal Growth Factor Stimulation

PloS One
Junfeng TongZhixiang Wang

Abstract

The small GTPase RhoA has been implicated in various cellular activities, including the formation of stress fibers, cell motility, and cytokinesis. In addition to the canonical GTPase cycle, recent findings have suggested that phosphorylation further contributes to the tight regulation of Rho GTPases. Indeed, RhoA is phosphorylated on serine 188 (188S) by a number of protein kinases. We have recently reported that Rac1 is phosphorylated on threonine 108 (108T) by extracellular signal-regulated kinases (ERK) in response to epidermal growth factor (EGF) stimulation. Here, we provide evidence that RhoA is phosphorylated by ERK on 88S and 100T in response to EGF stimulation. We show that ERK interacts with RhoA and that this interaction is dependent on the ERK docking site (D-site) at the C-terminus of RhoA. EGF stimulation enhanced the activation of the endogenous RhoA. The phosphomimetic mutant, GFP-RhoA S88E/T100E, when transiently expressed in COS-7 cells, displayed higher GTP-binding than wild type RhoA. Moreover, the expression of GFP-RhoA S88E/T100E increased actin stress fiber formation in COS-7 cells, which is consistent with its higher activity. In contrast to Rac1, phosphorylation of RhoA by ERK does not target RhoA to t...Continue Reading

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Citations

Aug 24, 2016·The Journal of Cell Biology·Thomas JungasAlice Davy
Jul 12, 2017·Small GTPases·Rebecca R FlorkeMichael J Hamann
Oct 19, 2017·International Journal of Molecular Sciences·Daniel Brandwein, Zhixiang Wang
Jan 30, 2018·Journal of Cellular Physiology·Jae-Gyu KimJae-Bong Park
Jun 1, 2018·International Journal of Molecular Sciences·Will LinthicumQi Wen
Jun 30, 2019·Cells·Claudia Voena, Roberto Chiarle
Dec 5, 2017·Small GTPases·Eun-Kyoung ChoiJae-Bong Park
Jun 18, 2020·Journal of Cell Science·Tejeshwar C RaoAlexa L Mattheyses
Sep 5, 2019·Cells·Abdalla AbdrabouZhixiang Wang
Jan 9, 2021·FASEB Journal : Official Publication of the Federation of American Societies for Experimental Biology·Abdalla Abdrabou, Zhixiang Wang
Jan 27, 2021·Cells·Zhong-Jian ShenJames S Malter
Jun 24, 2017·The American Journal of the Medical Sciences·Rui YamaguchiYasuo Yamaguchi
May 7, 2021·Biological & Pharmaceutical Bulletin·Kentaro YanoTakuo Ogihara
Aug 8, 2021·International Journal of Molecular Sciences·Nadezhda BarvitenkoAntonella Pantaleo

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Methods Mentioned

BETA
GTPases
GTPase
nucleotide exchange
transfection
pull-down
pull down
Co-immunoprecipitation
co-IP
pulled down
pulldowns

Software Mentioned

Northern Eclipse
SPSS
DeltaVision SoftWoRx
Delta Vision SoftWoRx
Empix

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