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Cited by 63Protein Kinase Regulation and GTPase SignalingCellular transport and secretionCellular Mechanics and Interactions

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

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Tong, Junfeng, Li, Laiji, Ballermann, Barbara, Wang, Zhixiang·2016·PLOS ONE
DOI 10.1371/journal.pone.0147103PMC PMC4729484
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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 the nucleus. Finally, we show that regardless of the phosphorylation status of RhoA and Rac1, substitution of the RhoA PBR with the Rac1 PBR targets RhoA to the nucleus and substitution of Rac1 PBR with RhoA PBR significantly reduces the nuclear localization of Rac1. In conclusion, ERK phosphorylates RhoA on 88S and 100T in response to EGF, which upregulates RhoA activity.

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  • ERK phosphorylates RhoA on Ser88 and Thr100, which upregulates RhoA activity as evaluated by its binding capacity to Rhotekin.

    “Extracellular signal-related kinase (ERK) phosphorylates RhoA on Ser88 and Thr100, which has been reported to upregulate RhoA activity as evaluated by its binding capacity to the effector protein Rhotekin [61].”

    From RhoA Signaling in Neurodegenerative Diseases (2022) · cites this paper as [61]

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Authors
Tong, J., Li, L., Ballermann, B., & Wang, Z.
Affiliations
University of Alberta (CA)
Corresponding author
Wang, Zhixiang
ORCID
Venue
PLOS ONE
Publisher
Public Library of Science (PLoS)
Published
2016-01-27
Volume
11
Issue
1
Pages
e0147103
ISSN
1932-6203 (ISSN-L)
DOI
10.1371/journal.pone.0147103
PMC
PMC4729484
OpenAlex
W2412949340
Cited by
63
References
1
Citations by year
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FWCI
2.42
Type
journal article
Language
en
Open access
Licence
Locations
  • PLoS ONE (publishedVersion, cc-by)
  • PubMed Central (publishedVersion, cc-by)
  • PubMed (publishedVersion)
  • DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion, cc-by-sa)
Funders
  • Alberta Cancer Foundation
  • University of Alberta
  • School of Medicine, New York University
  • Canadian Institutes of Health Research (RES0006924)
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Protein Kinase Regulation and GTPase Signaling
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Cell Biology › Cellular transport and secretion
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Cell Biology › Cellular Mechanics and Interactions
Keywords
RhoA, ERK, epidermal growth factor, RhoA activation, small GTPase regulation, Rac1, stress fiber formation, nuclear localization, phosphomimetic mutant, cell motility
MeSH terms
Actins, Animals, Cell Line, Cell Nucleus, Chlorocebus aethiops, Cytoskeleton, Epidermal Growth Factor, Humans, Phosphorylation, Signal Transduction, COS Cells, rhoA GTP-Binding Protein, Extracellular Signal-Regulated MAP Kinases

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