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In vivo assembly of a Dictyostelium lamin mutant induced by light, mechanical stress, and pH

  • We expressed Dictyostelium lamin (NE81) lacking both a functional nuclear localization signal and a CAAX-box for C-terminal lipid modification. This lamin mutant assembled into supramolecular, three-dimensional clusters in the cytosol that disassembled at the onset of mitosis and re-assembled in late telophase, thus mimicking the behavior of the endogenous protein. As disassembly is regulated by CDK1-mediated phosphorylation at serine 122, we generated a phosphomimetic S122E mutant called GFP-NE81-S122E-∆NLS∆CLIM. Surprisingly, during imaging, the fusion protein assembled into cytosolic clusters, similar to the protein lacking the phosphomimetic mutation. Clusters disassembled again in the darkness. Assembly could be induced with blue but not green or near ultraviolet light, and it was independent of the fusion tag. Assembly similarly occurred upon cell flattening. Earlier reports and own observations suggested that both blue light and cell flattening could result in a decrease of intracellular pH. Indeed, keeping the cells at low pHWe expressed Dictyostelium lamin (NE81) lacking both a functional nuclear localization signal and a CAAX-box for C-terminal lipid modification. This lamin mutant assembled into supramolecular, three-dimensional clusters in the cytosol that disassembled at the onset of mitosis and re-assembled in late telophase, thus mimicking the behavior of the endogenous protein. As disassembly is regulated by CDK1-mediated phosphorylation at serine 122, we generated a phosphomimetic S122E mutant called GFP-NE81-S122E-∆NLS∆CLIM. Surprisingly, during imaging, the fusion protein assembled into cytosolic clusters, similar to the protein lacking the phosphomimetic mutation. Clusters disassembled again in the darkness. Assembly could be induced with blue but not green or near ultraviolet light, and it was independent of the fusion tag. Assembly similarly occurred upon cell flattening. Earlier reports and own observations suggested that both blue light and cell flattening could result in a decrease of intracellular pH. Indeed, keeping the cells at low pH also reversibly induced cluster formation. Our results indicate that lamin assembly can be induced by various stress factors and that these are transduced via intracellular acidification. Although these effects have been shown in a phosphomimetic CDK1 mutant of the Dictyostelium lamin, they are likely relevant also for wild-type lamin.zeige mehrzeige weniger

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Metadaten
Verfasserangaben:Marianne GrafeGND, Phillip Hofmann, Petros BatsiosORCiDGND, Irene Meyer, Ralph GräfORCiDGND
URN:urn:nbn:de:kobv:517-opus4-525075
DOI:https://doi.org/10.25932/publishup-52507
ISSN:1866-8372
Titel des übergeordneten Werks (Deutsch):Postprints der Universität Potsdam : Mathematisch-Naturwissenschaftliche Reihe
Schriftenreihe (Bandnummer):Zweitveröffentlichungen der Universität Potsdam : Mathematisch-Naturwissenschaftliche Reihe (1213)
Publikationstyp:Postprint
Sprache:Englisch
Datum der Erstveröffentlichung:23.04.2020
Erscheinungsjahr:2020
Veröffentlichende Institution:Universität Potsdam
Datum der Freischaltung:19.11.2021
Freies Schlagwort / Tag:NE81; lamin; nuclear envelope; nuclear lamina
Dictyostelium
Ausgabe:8
Aufsatznummer:1834
Seitenanzahl:16
Quelle:Cells 2020, 9(8), 1834; https://doi.org/10.3390/cells9081834
Organisationseinheiten:Mathematisch-Naturwissenschaftliche Fakultät / Institut für Biochemie und Biologie
DDC-Klassifikation:5 Naturwissenschaften und Mathematik / 57 Biowissenschaften; Biologie / 570 Biowissenschaften; Biologie
Peer Review:Referiert
Publikationsweg:Open Access / Green Open-Access
Lizenz (Deutsch):License LogoCC-BY - Namensnennung 4.0 International
Externe Anmerkung:Bibliographieeintrag der Originalveröffentlichung/Quelle
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