Extracellular matrix controls tubulin monomer levels in hepatocytes by regulating protein turnover.
Open accessAbstract
Cells have evolved an autoregulatory mechanism to dampen variations in the concentration of tubulin monomer that is available to polymerize into microtubules (MTs), a process that is known as tubulin autoregulation. However, thermodynamic analysis of MT polymerization predicts that the concentration of free tubulin monomer must vary if MTs are to remain stable under different mechanical loads that result from changes in cell adhesion to the extracellular matrix (ECM). To determine how these seemingly contradictory regulatory mechanisms coexist in cells, we measured changes in the masses of tubulin monomer and polymer that resulted from altering cell-ECM contacts. Primary rat hepatocytes were cultured in chemically defined medium on bacteriological petri dishes that were precoated with different densities of laminin (LM). Increasing the LM density from low to high (1-1000 ng/cm2), promoted cell spreading (average projected cell area increased from 1200 to 6000 microns2) and resulted in formation of a greatly extended MT network. Nevertheless, the steady-state mass of tubulin polymer was similar at 48 h, regardless of cell shape or ECM density. In contrast, round hepatocytes on low LM contained a threefold higher mass of tubulin monomer when compared with spread cells on high LM. Furthermore, similar results were obtained whether LM, fibronectin, or type I collagen were used for cell attachment. Tubulin autoregulation appeared to function normally in these cells because tubulin mRNA levels and protein synthetic rates were greatly depressed in round cells that contained the highest level of free tubulin monomer. However, the rate of tubulin protein degradation slowed, causing the tubulin half-life to increase from approximately 24 to 55 h as the LM density was lowered from high to low and cell rounding was promoted. These results indicate that the set-point for the tubulin monomer mass in hepatocytes can be regulated by altering the density of ECM contacts and changing cell shape. This finding is consistent with a mechanism of MT regulation in which the ECM stabilizes MTs by both accepting transfer of mechanical loads and altering tubulin degradation in cells that continue to autoregulate tubulin synthesis.
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Increased percentages of depolymerized tubulin in cells treated with TTFields while spreading following re-plating supports possibility that TTFields interfere with polymerization of microtubules.
“Increased percentages of depolymerized tubulin in cells treated with TTFields while spreading following re-plating supports the possibility that TTFields interfere with polymerization of microtubules42.”
From Mitotic Spindle Disruption by Alternating Electric Fields Leads to Improper Chromosome Segregation and Mitotic Catastrophe in Cancer Cells (2015) · cites this paper as [42]
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- Authors
- Mooney, D. J., Hansen, L. K., Langer, R., Vacanti, J. P., & Ingber, D. E.
- Affiliations
- Massachusetts Institute of Technology (US)
- Venue
- Molecular Biology of the Cell
- Published
- 1994-12
- Volume
- 5
- Issue
- 12
- Pages
- 1281-1288
- ISSN
- 1059-1524 (ISSN-L)
- 1939-4586
- PMC
- PMC301157
- OpenAlex
- W1972983450
- Cited by
- 53
- References
- 34
- Citations by year
- 2026: 1 · 2020: 2 · 2018: 1 · 2017: 1 · 2016: 3 · 2015: 2 · 2014: 1 · 2013: 3 · 2012: 2
- FWCI
- 0.83
- Type
- journal article
- Language
- en
- Open access
- green
- www.ncbi.nlm.nih.gov
- Locations
- Molecular Biology of the Cell (publishedVersion)
- Europe PMC (PubMed Central) (publishedVersion)
- PubMed Central (publishedVersion)
- PubMed (publishedVersion)
- Funders
- American Cancer Society
- School of Medicine, Boston University
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Cell Biology › Microtubule and mitosis dynamics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Cell Biology › Cellular Mechanics and Interactions
- Health Sciences › Medicine › Immunology and Allergy › Cell Adhesion Molecules Research
- Keywords
- hepatocytes, extracellular matrix, laminin, cell adhesion, cell shape, microtubule stability, tubulin synthesis, cell spreading, fibronectin, type I collagen
- MeSH terms
- Animals, Cell Movement, Cells, Cultured, Extracellular Matrix, Homeostasis, Liver, Male, Tubulin, Rats, Wistar, Rats
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