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Cited by 30Cellular Mechanics and InteractionsProtein Kinase Regulation and GTPase SignalingHippo pathway signaling and YAP/TAZ

Compressive Stress Induces Dephosphorylation of the Myosin Regulatory Light Chain via RhoA Phosphorylation by the Adenylyl Cyclase/Protein Kinase A Signaling Pathway

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Takemoto, Kenji, Ishihara, Seiichiro, Mizutani, Takeomi, Kawabata, Kazushige, Haga, Hisashi·2015·PLOS ONE
DOI 10.1371/journal.pone.0117937PMC PMC4348516
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Abstract

Mechanical stress that arises due to deformation of the extracellular matrix (ECM) either stretches or compresses cells. The cellular response to stretching has been actively studied. For example, stretching induces phosphorylation of the myosin regulatory light chain (MRLC) via the RhoA/RhoA-associated protein kinase (ROCK) pathway, resulting in increased cellular tension. In contrast, the effects of compressive stress on cellular functions are not fully resolved. The mechanisms for sensing and differentially responding to stretching and compressive stress are not known. To address these questions, we investigated whether phosphorylation levels of MRLC were affected by compressive stress. Contrary to the response in stretching cells, MRLC was dephosphorylated 5 min after cells were subjected to compressive stress. Compressive loading induced activation of myosin phosphatase mediated via the dephosphorylation of myosin phosphatase targeting subunit 1 (Thr853). Because myosin phosphatase targeting subunit 1 (Thr853) is phosphorylated only by ROCK, compressive loading may have induced inactivation of ROCK. However, GTP-bound RhoA (active form) increased in response to compressive stress. The compression-induced activation of RhoA and inactivation of its effector ROCK are contradictory. This inconsistency was due to phosphorylation of RhoA (Ser188) that reduced affinity of RhoA to ROCK. Treatment with the inhibitor of protein kinase A that phosphorylates RhoA (Ser188) induced suppression of compression-stimulated MRLC dephosphorylation. Incidentally, stretching induced phosphorylation of MRLC, but did not affect phosphorylation levels of RhoA (Ser188). Together, our results suggest that RhoA phosphorylation is an important process for MRLC dephosphorylation by compressive loading, and for distinguishing between stretching and compressing cells.

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  • Ser188 phosphorylation inhibits RhoA binding to its effector ROCK but does not affect binding to rhotekin and mDia, thereby specifically inhibiting RhoA/ROCK signaling.

    “Furthermore, Ser188 phosphorylation has been found to inhibit RhoA binding to its effector protein Rho-associated coiled-coil-containing kinase (ROCK) but does not affect its binding to other RhoA effector proteins, including rhotekin and mammalian diaphanous (mDia), thereby specifically inhibiting RhoA/ROCK signaling [53,54].”

    From RhoA Signaling in Neurodegenerative Diseases (2022) · cites this paper as [53]

    The Results section on RhoA activation and phosphorylation (page not provided) says Ser188-phosphorylated RhoA cannot interact with or activate ROCK. The paper uses rhotekin to assay active RhoA and discusses mDia1 signaling, but it does not establish that Ser188 phosphorylation leaves binding to either effector unaffected. Thus the claimed selectivity for ROCK over those effectors is not supported. (LLM, conf=0.86)

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Authors
Takemoto, K., Ishihara, S., Mizutani, T., Kawabata, K., & Haga, H.
Affiliations
Hokkaido University (JP)
Corresponding author
Haga, Hisashi
ORCID
Venue
PLOS ONE
Publisher
Public Library of Science (PLoS)
Published
2015-03-03
Volume
10
Issue
3
Pages
e0117937
ISSN
1932-6203 (ISSN-L)
DOI
10.1371/journal.pone.0117937
PMC
PMC4348516
OpenAlex
W2058490656
Cited by
30
References
0
Citations by year
2026: 1 · 2025: 2 · 2024: 2 · 2023: 2 · 2022: 5 · 2021: 5 · 2020: 3 · 2019: 3 · 2018: 1 · 2017: 1 · 2016: 3 · 2015: 2
FWCI
1.12
Type
journal article
Language
en
Open access
Licence
Locations
  • PLoS ONE (publishedVersion, cc-by)
  • Europe PMC (PubMed Central) (publishedVersion, cc-by)
  • PubMed Central (publishedVersion, cc-by)
  • PubMed (publishedVersion)
  • Institutional Repositories DataBase (IRDB) (submittedVersion, cc-by)
  • DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion, cc-by-sa)
  • Hokkaido University Collection of Scholarly and Academic Papers (Hokkaido University) (submittedVersion, cc-by)
  • Figshare (submittedVersion, cc-by)
Funders
  • Ministry of Education, Culture, Sports, Science and Technology (26430104, 26106704, 25287106)
  • Japan Society for the Promotion of Science (25287106, 26430104, 24370069)
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Cell Biology › Cellular Mechanics and Interactions
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Protein Kinase Regulation and GTPase Signaling
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Cell Biology › Hippo pathway signaling and YAP/TAZ
Keywords
compressive stress, myosin regulatory light chain, myosin phosphatase, extracellular matrix deformation, cell stretching, mechanotransduction
MeSH terms
Adenylyl Cyclases, Animals, Cell Line, Marine Toxins, Actin Cytoskeleton, Myosin-Light-Chain Kinase, Oxazoles, Phosphorylation, Signal Transduction, Cyclic AMP-Dependent Protein Kinases, Myosin Light Chains, Compressive Strength, rhoA GTP-Binding Protein, RNA Interference, RNA, Small Interfering, Myosin-Light-Chain Phosphatase, Mice, rho-Associated Kinases

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