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Cited by 55Protein Kinase Regulation and GTPase SignalingIon Transport and Channel RegulationCellular Mechanics and Interactions

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Depolarization induces Rho-Rho kinase-mediated myosin light chain phosphorylation in kidney tubular cells

Szászi, Katalin, Sirokmány, Gábor, Ciano-Oliveira, Caterina Di, Rotstein, Ori D., Kapus, András·2005·American Journal of Physiology-Cell Physiology
DOI 10.1152/ajpcell.00481.2004
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Abstract

Myosin-based contractility plays important roles in the regulation of epithelial functions, particularly paracellular permeability. However, the triggering factors and the signaling pathways that control epithelial myosin light chain (MLC) phosphorylation have not been elucidated. Herein we show that plasma membrane depolarization provoked by distinct means, including high extracellular K(+), the lipophilic cation tetraphenylphosphonium, or the ionophore nystatin, induced strong diphosphorylation of MLC in kidney epithelial cells. In sharp contrast to smooth muscle, depolarization of epithelial cells did not provoke a Ca(2+) signal, and removal of external Ca(2+) promoted rather than inhibited MLC phosphorylation. Moreover, elevation of intracellular Ca(2+) did not induce significant MLC phosphorylation, and the myosin light chain kinase (MLCK) inhibitor ML-7 did not prevent the depolarization-induced MLC response, suggesting that MLCK is not a regulated element in this process. Instead, the Rho-Rho kinase (ROK) pathway is the key mediator because 1) depolarization stimulated Rho and induced its peripheral translocation, 2) inhibition of Rho by Clostridium difficile toxin B or C3 transferase abolished MLC phosphorylation, and 3) the ROK inhibitor Y-27632 suppressed the effect. Importantly, physiological depolarizing stimuli were able to activate the same pathway: L-alanine, the substrate of the electrogenic Na(+)-alanine cotransporter, stimulated Rho and induced Y-27632-sensi…

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Metadata

Authors
Szászi, K., Sirokmány, G., Ciano-Oliveira, C. D., Rotstein, O. D., & Kapus, A.
Affiliations
University Health Network (CA)
ORCID
Venue
American Journal of Physiology-Cell Physiology
Publisher
American Physiological Society
Published
2005-09
Volume
289
Issue
3
Pages
C673-C685
ISSN
  • 0363-6143 (ISSN-L)
  • 1522-1563
DOI
10.1152/ajpcell.00481.2004
OpenAlex
W2036138598
Cited by
55
References
0
Citations by year
2026: 1 · 2025: 1 · 2024: 2 · 2021: 2 · 2020: 1 · 2019: 4 · 2017: 1 · 2016: 1 · 2015: 1 · 2014: 3 · 2013: 4 · 2012: 6
FWCI
1.76
Type
journal article
Language
en
Open access
closed
Locations
  • American Journal of Physiology-Cell Physiology (publishedVersion)
  • PubMed (publishedVersion)
Funders
Canadian Institutes of Health Research (63790)
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Protein Kinase Regulation and GTPase Signaling
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Ion Transport and Channel Regulation
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Cell Biology › Cellular Mechanics and Interactions
Keywords
myosin light chain phosphorylation, plasma membrane depolarization, Rho kinase, RhoA, renal tubular epithelial cells, paracellular permeability, L-alanine, tetraphenylphosphonium, nystatin, calcium-independent signaling, myosin light chain kinase, Y-27632, Clostridium difficile toxin B, C3 transferase
MeSH terms
Animals, Calcium, Cell Membrane, Dogs, Electric Stimulation, Epithelial Cells, Kidney Tubules, Distal, Membrane Potentials, Phosphorylation, Potassium, Swine, Protein Serine-Threonine Kinases, LLC-PK1 Cells, Myosin Light Chains, rho GTP-Binding Proteins, Amino Acid Transport Systems, Neutral, Symporters, Intracellular Signaling Peptides and Proteins, rho-Associated Kinases

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