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PMID: 3611184 Published · ppublish English Comparative Study Journal Article Research Support, Non-U.S. Gov't

Exocytosis induction in Paramecium tetraurelia cells by exogenous phosphoprotein phosphatase in vivo and in vitro: possible involvement of calcineurin in exocytotic membrane fusion.

The Journal of cell biology ·Vol. 105 ·No. 1 ·1987-07-00 ·Pages 181-9

Momayezi M, Lumpert CJ, Kersken H, Gras U, Plattner H, Krinks MH, Klee CB

Abstract

Since it had been previously shown that in Paramecium cells exocytosis involves the dephosphorylation of a 65-kD phosphoprotein (PP), we tried to induce exocytotic membrane fusion by exogenous phosphatases (alkaline phosphatase or calcineurin [CaN]). The occurrence of calmodulin (CaM) at preformed exocytosis sites (Momayezi, M., H. Kersken, U. Gras, J. Vilmart-Seuwen, and H. Plattner, 1986, J. Histochem. Cytochem., 34:1621-1638) and the current finding of the presence of the 65-kD PP and of a CaN-like protein in cell surface fragments ("cortices") isolated from Paramecium cells led us to also test the effect of antibodies (Ab) against CaM or CaN on exocytosis performance. Microinjected anti-CaN Ab strongly inhibit exocytosis. (Negative results with microinjected anti-CaM Ab can easily be explained by the abundance of CaM.) Alternatively, microinjection of a Ca2+-CaM-CaN complex triggers exocytosis. The same occurs with alkaline phosphatase. All these effects can also be mimicked in vitro with isolated cortices. In vitro exocytosis triggered by adding Ca2+-CaM-CaN or alkaline phosphatase is paralleled by dephosphorylation of the 65-kD PP. Exocytosis can also be inhibited in cortices by anti-CaM Ab or anti-CaN Ab. In wild-type cells, compounds that inhibit phosphatase activity, but none that inhibit kinases or proteases, are able to inhibit exocytosis. Exocytosis cannot be induced by phosphatase injection in a membrane-fusion-deficient mutant strain (nd9-28 degrees C) characterized by a defective organization of exocytosis sites (Beisson, J., M. Lefort-Tran, M. Pouphile, M. Rossignol, and B. Satir, 1976, J. Cell Biol., 69:126-143). We conclude that exocytotic membrane fusion requires an adequate assembly of molecular components to allow for the dephosphorylation of a 65-kD PP and that this step is crucial for the induction of exocytotic membrane fusion in Paramecium cells. In vivo this probably involves a Ca2+-CaM-stimulated CaN-like PP phosphatase.

MeSH Terms
Alkaline Phosphatase/antagonists & inhibitors,pharmacology Animals Antibodies/immunology Calmodulin/immunology,metabolism Calmodulin-Binding Proteins/immunology,pharmacology,physiology Exocytosis Membrane Fusion Microinjections Paramecium/drug effects,physiology Phosphoproteins/metabolism Phosphorylation
Chemicals
Antibodies Calmodulin Calmodulin-Binding Proteins Phosphoproteins Alkaline Phosphatase
Authors & Affiliations
7 authors, click to expand affiliations / ORCID
Momayezi M
Lumpert C J
Kersken H
Gras U
Plattner H
Krinks M H
Klee C B
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Article Info
Journal
The Journal of cell biology
Abbr.
J Cell Biol
ISSN
0021-9525
DOI
10.1083/jcb.105.1.181
Published
1987-07-00
Pages
181-9
Language
English
Region
United States
NLM ID
0375356
PMCID
PMC2114937
Subset
IM
External Links
PubMed source DOI: 10.1083/jcb.105.1.181 PMC Full Text View journal details
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