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PMID: 38946197 Published · ppublish English Journal Article

RhoB plays a central role in hyperosmolarity-induced cell shrinkage in renal cells.

Journal of cellular physiology ·Vol. 239 ·No. 9 ·2024-09-00 ·Pages e31343

Centrone M, Saltarella I, D'Agostino M, Ranieri M, Venneri M, Di Mise A, Simone L, Pisani F, Valenti G, Frassanito MA, Tamma G

Abstract

The small Rho GTP-binding proteins are important cell morphology, function, and apoptosis regulators. Unlike other Rho proteins, RhoB can be subjected to either geranylgeranylation (RhoB-GG) or farnesylation (RhoB-F), making that the only target of the farnesyltransferase inhibitor (FTI). Fluorescence resonance energy transfer experiments revealed that RhoB is activated by hyperosmolarity. By contrast, hyposmolarity did not affect RhoB activity. Interestingly, treatment with farnesyltransferase inhibitor-277 (FTI-277) decreased the cell size. To evaluate whether RhoB plays a role in volume reduction, renal collecting duct MCD4 cells and Human Kidney, HK-2 were transiently transfected with RhoB-wildtype-Enhance Green Fluorescence Protein (RhoB-wt-EGFP) and RhoB-CLLL-EGFP which cannot undergo farnesylation. A calcein-based fluorescent assay revealed that hyperosmolarity caused a significant reduction of cell volume in mock and RhoB-wt-EGFP-expressing cells. By contrast, cells treated with FTI-277 or expressing the RhoB-CLLL-EGFP mutant did not properly respond to hyperosmolarity with respect to mock and RhoB-wt-EGFP expressing cells. These findings were further confirmed by 3D-LSCM showing that RhoB-CLLL-EGFP cells displayed a significant reduction in cell size compared to cells expressing RhoB-wt-EGFP. Moreover, flow cytometry analysis revealed that RhoB-CLLL-EGFP expressing cells as well as FTI-277-treated cells showed a significant increase in cell apoptosis. Together, these data suggested that: (i) RhoB is sensitive to hyperosmolarity and not to hyposmolarity; (ii) inhibition of RhoB farnesylation associates with an increase in cell apoptosis, likely suggesting that RhoB might be a paramount player controlling apoptosis by interfering with responses to cell volume change.

Keywords
Rho GTP‐binding proteins RhoB cell volume hyperosmolarity kidney
MeSH Terms
Cell Size/drug effects Kidney/cytology,metabolism Osmosis Humans Animals Mice Farnesyltranstransferase/antagonists & inhibitors Enzyme Inhibitors/pharmacology rhoB GTP-Binding Protein/genetics,metabolism Apoptosis/drug effects Mutation
Chemicals
Farnesyltranstransferase Enzyme Inhibitors rhoB GTP-Binding Protein
Authors & Affiliations
11 authors, click to expand affiliations / ORCID
Centrone Mariangela
Department of Biosciences Biotechnologies and Environment, University of Bari Aldo Moro, Bari, Italy.
Saltarella Ilaria
Department of Precision and Regenerative Medicine and Ionian Area, Section of Pharmacology, School of Medicine, University of Bari Aldo Moro, Bari, Italy.
D'Agostino Mariagrazia
Department of Biosciences Biotechnologies and Environment, University of Bari Aldo Moro, Bari, Italy.
Ranieri Marianna
Department of Biosciences Biotechnologies and Environment, University of Bari Aldo Moro, Bari, Italy.
Venneri Maria
Istituti Clinici Scientifici Maugeri SPA SB IRCCS, Bari, Italy.
Di Mise Annarita
Department of Biosciences Biotechnologies and Environment, University of Bari Aldo Moro, Bari, Italy.
Simone Laura
Fondazione IRCCS Casa Sollievo della Sofferenza, Cancer Stem Cells Unit, San Giovanni Rotondo, Italy.
Pisani Francesco
Department of Biosciences Biotechnologies and Environment, University of Bari Aldo Moro, Bari, Italy.
Valenti Giovanna
Department of Biosciences Biotechnologies and Environment, University of Bari Aldo Moro, Bari, Italy.
Frassanito Maria A
Department of Precision and Regenerative Medicine and Ionian Area, Section of Clinical Pathology, University of Bari Aldo Moro, Bari, Italy.
Tamma Grazia ORCID
Department of Biosciences Biotechnologies and Environment, University of Bari Aldo Moro, Bari, Italy.
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Article Info
Journal
Journal of cellular physiology
Abbr.
J Cell Physiol
ISSN
1097-4652
DOI
10.1002/jcp.31343
Published
2024-09-00
Epub
2024-00-30
Pages
e31343
Language
English
Region
United States
NLM ID
0050222
Subset
IM
Grants
Ministero dell'Università e della Ricerca
External Links
PubMed source DOI: 10.1002/jcp.31343 View journal details
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