Cell cycle-dependent dynamics of a plant intermediate filament motif protein with intracellular localization related to
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ORIGINAL ARTICLE
Cell cycle-dependent dynamics of a plant intermediate filament motif protein with intracellular localization related to microtubules Hikaru Utsunomiya 1 & Masayuki Fujita 1 & Fumio Naito 1 & Tsuyoshi Kaneta 1 Received: 6 November 2019 / Accepted: 7 May 2020 # Springer-Verlag GmbH Austria, part of Springer Nature 2020
Abstract Although intermediate filaments (IFs) are biochemically and immunologically suggested to exist in plant cells, there are few molecular genetic studies related to the proteins that form these structures. In this study, Arabidopsis AT3G05270 was selected as a candidate gene for a protein constituting IF in plant cells. The protein encoded by AT3G05270 has a large α-helix as well as the IF protein motif indispensable for maintaining the structures of IF. Moreover, fluorescence signals of this protein fused with GFP exhibited cytoskeleton-like filamentous structures in plant cells. Thus, we named the protein encoded by AT3G05270 as Intermediate Filament Motif Protein 1 (IFMoP1). The structures composed of IFMoP1 and their localizations were examined in IFMoP1-GFP-expressing tobacco BY-2 cells whose cell cycle was synchronized using aphidicolin, a DNA synthesis inhibitor, and propyzamide, a microtubule-disrupting agent. The IFMoP1-GFP signals were present at the spindles and phragmoplasts in the mitotic phase. In addition, the frequency of cells with cytoskeleton-like filamentous structures composed of IFMoP1-GFP increased with the increase in cells that completed cell division, and then decreased after several hours. In terms of the relationship in intracellular localization between IFMoP1 and microtubules, the filamentous structures composed of IFMoP1 were present independently of microtubules during interphase. In living cells, these filamentous structures moved along with the nucleus. IFMoP1 co-localized with spindle and phragmoplast microtubules during mitosis, as well as with a part of the cortical microtubules in interphase. Keywords Intermediate filament . Microtubules . Arabidopsis . Tobacco BY-2 cells
Introduction Intermediate filaments (IFs) are cytoskeletal protein filaments of 10 nm in diameter with superior flexibility and mechanical strength. In animal cells, IFs provide mechanical stability and maintain cell and tissue morphology. Many different genes encoding the IF proteins have been identified in animals (Minin and Moldaver 2008). While, in plant cells, the existence of IFs has not been definitively established. However, Handling Editor: Anne-Catherine Schmit Electronic supplementary material The online version of this article (https://doi.org/10.1007/s00709-020-01512-1) contains supplementary material, which is available to authorized users. * Tsuyoshi Kaneta [email protected] 1
Department of Chemistry & Biology, Graduate School of Science & Engineering, Ehime University, 2-5 Bunkyo-cho, Matsuyama 790-8577, Japan
there is biochemical and immunological evidence that indicates the presence of IFs in plant cells. Fibrillar bundles composed of fi
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