Here we describe the phenotypic characterization of the cta4+ gene, encoding a novel member of the P4 family of P-type ATPases of fission yeast. The cta4Δ mutant is temperature sensitive and cold sensitive lethal and displays several morphological defects in cell polarity and cytokinesis. Microtubules are generally destabilized in cells lacking Cta4p. The microtubule length is decreased, and the number of microtubules per cell is increased. This is concomitant with an increase in the number of microtubule catastrophe events in the midzone of the cell. These defects are likely due to a general imbalance in cation homeostasis. Immunofluorescence microscopy and membrane fractionation experiments revealed that green fluorescent protein–tagged Cta4 localizes to the ER. Fluorescence resonance energy transfer experiments in living cells using the yellow cameleon indicator for Ca2+ indicated that Cta4p regulates the cellular Ca2+ concentration. Thus, our results reveal a link between cation homeostasis and the control of cell shape, microtubule dynamics, and cytokinesis, and appoint Ca2+ as a key ion in controlling these processes.
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10 June 2002
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June 10 2002
The endoplasmic reticulum cation P-type ATPase Cta4p is required for control of cell shape and microtubule dynamics
Anna L. Okorokova Façanha,
Anna L. Okorokova Façanha
1Laboratório Fisiologia e Bioquímica de Microrganismos, Centro de Biociências e Biotecnologia, Universidade Estadual do Norte Fluminense, Campos dos Goytacazes, Horto, CEP 28015-620, Brazil
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Henrik Appelgren,
Henrik Appelgren
2Karolinska Institutet, Department of Biosciences Novum/University College Sodertorn, Deptartment of Natural Sciences, S-141 04 Huddinge, Sweden
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Mohammad Tabish,
Mohammad Tabish
2Karolinska Institutet, Department of Biosciences Novum/University College Sodertorn, Deptartment of Natural Sciences, S-141 04 Huddinge, Sweden
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Lev Okorokov,
Lev Okorokov
1Laboratório Fisiologia e Bioquímica de Microrganismos, Centro de Biociências e Biotecnologia, Universidade Estadual do Norte Fluminense, Campos dos Goytacazes, Horto, CEP 28015-620, Brazil
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Karl Ekwall
Karl Ekwall
2Karolinska Institutet, Department of Biosciences Novum/University College Sodertorn, Deptartment of Natural Sciences, S-141 04 Huddinge, Sweden
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Anna L. Okorokova Façanha
1Laboratório Fisiologia e Bioquímica de Microrganismos, Centro de Biociências e Biotecnologia, Universidade Estadual do Norte Fluminense, Campos dos Goytacazes, Horto, CEP 28015-620, Brazil
Henrik Appelgren
2Karolinska Institutet, Department of Biosciences Novum/University College Sodertorn, Deptartment of Natural Sciences, S-141 04 Huddinge, Sweden
Mohammad Tabish
2Karolinska Institutet, Department of Biosciences Novum/University College Sodertorn, Deptartment of Natural Sciences, S-141 04 Huddinge, Sweden
Lev Okorokov
1Laboratório Fisiologia e Bioquímica de Microrganismos, Centro de Biociências e Biotecnologia, Universidade Estadual do Norte Fluminense, Campos dos Goytacazes, Horto, CEP 28015-620, Brazil
Karl Ekwall
2Karolinska Institutet, Department of Biosciences Novum/University College Sodertorn, Deptartment of Natural Sciences, S-141 04 Huddinge, Sweden
Address correspondence to Karl Ekwall, University College Sodertorn, Dept. of Natural Sciences, Alfred Nobels Alle 3, S-141 52 Huddinge, Sweden. Tel.: 46-8-608-4713. Fax: 46-8-608-4510. E-mail: [email protected]
*
Abbreviations used in this paper: CaM, calmodulin; CoA, coenzyme A; CsA, cyclosporin A; CYP, cyan fluorescent protein; FRET, fluorescence resonance energy transfer; GFP, green fluorescent protein; IF, immunofluorescence; SERCA, sarco/ER Ca2+-ATPases; TBZ, thiabendazole; TMD, transmembrane spanning domain; YFP, yellow fluorescent protein.
Received:
November 05 2001
Revision Received:
February 28 2002
Accepted:
April 03 2002
Online ISSN: 1540-8140
Print ISSN: 0021-9525
The Rockefeller University Press
2002
J Cell Biol (2002) 157 (6): 1029–1040.
Article history
Received:
November 05 2001
Revision Received:
February 28 2002
Accepted:
April 03 2002
Citation
Anna L. Okorokova Façanha, Henrik Appelgren, Mohammad Tabish, Lev Okorokov, Karl Ekwall; The endoplasmic reticulum cation P-type ATPase Cta4p is required for control of cell shape and microtubule dynamics . J Cell Biol 10 June 2002; 157 (6): 1029–1040. doi: https://doi.org/10.1083/jcb.200111012
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