Cortical cytoskeleton dynamics regulates plasma membrane calcium ATPase isoform-2 (PMCA2) activity.

Dalghi Gens, Marianela Gisela; Ferreira-Gomes, Mariela; Montalbetti, Nicolas; Simonin, Alexandre; Strehler, Emanuel E; Hediger, Matthias; Rossi, Juan Pablo (2017). Cortical cytoskeleton dynamics regulates plasma membrane calcium ATPase isoform-2 (PMCA2) activity. Biochimica et biophysica acta - molecular cell research, 1864(8), pp. 1413-1424. Elsevier 10.1016/j.bbamcr.2017.05.014

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We have previously shown that purified actin can directly bind to human plasma membrane Ca(2+) ATPase 4b (hPMCA4b) and exert a dual modulation on its Ca(2+)-ATPase activity: F-actin inhibits PMCA while short actin oligomers may contribute to PMCA activation. These studies had to be performed with purified proteins given the nature of the biophysical and biochemical approaches used. To assess whether a functional interaction between the PMCAs and the cortical cytoskeleton is of physiological relevance, we characterized this phenomenon in the context of a living cell by monitoring in real-time the changes in the cytosolic calcium levels ([Ca(2+)]CYT). In this study, we tested the influence of drugs that change the actin and microtubule polymerization state on the activity and membrane expression of the PMCA transiently expressed in human embryonic kidney (HEK293) cells, which allowed us to observe and quantify these relationships in a live cell, for the first time. We found that disrupting the actin cytoskeleton with cytochalasin D significantly increased PMCA-mediated Ca(2+) extrusion (~50-100%) whereas pre-treatment with the F-actin stabilizing agent jasplakinolide caused its full inhibition. When the microtubule network was disrupted by pretreatment of the cells with colchicine, we observed a significant decrease in PMCA activity (~40-60% inhibition) in agreement with the previously reported role of acetylated tubulin on the calcium pump. In none of these cases was there a difference in the level of expression of the pump at the cell surface, thus suggesting that the specific activity of the pump was the regulated parameter. Our results indicate that PMCA activity is profoundly affected by the polymerization state of the cortical cytoskeleton in living cells.

Item Type:

Journal Article (Original Article)

Division/Institute:

04 Faculty of Medicine > Pre-clinic Human Medicine > Institute of Biochemistry and Molecular Medicine

UniBE Contributor:

Dalghi Gens, Marianela Gisela; Montalbetti, Nicolas; Simonin, Alexandre and Hediger, Matthias

Subjects:

500 Science > 570 Life sciences; biology
600 Technology > 610 Medicine & health

ISSN:

0167-4889

Publisher:

Elsevier

Language:

English

Submitter:

Kevin Marc Rupp

Date Deposited:

20 Nov 2017 17:22

Last Modified:

26 Oct 2019 16:38

Publisher DOI:

10.1016/j.bbamcr.2017.05.014

PubMed ID:

28527708

Uncontrolled Keywords:

Actin Cortical cytoskeleton Microtubules Modulation PMCA2

BORIS DOI:

10.7892/boris.106122

URI:

https://boris.unibe.ch/id/eprint/106122

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