Gullmets Josef, Torvaldson Elin, Lindqvist Julia, Imanishi Susumu Y, Taimen Pekka, Meinander Annika, Eriksson John E
Cell Biology, Faculty of Science and Engineering, Åbo Akademi University, Turku, Finland.
Turku Centre for Biotechnology, University of Turku and Åbo Akademi University, Turku, Finland.
FASEB J. 2017 Dec;31(12):5332-5341. doi: 10.1096/fj.201700332R. Epub 2017 Aug 4.
Cytoplasmic intermediate filaments (cIFs) are found in all eumetazoans, except arthropods. To investigate the compatibility of cIFs in arthropods, we expressed human vimentin (hVim), a cIF with filament-forming capacity in vertebrate cells and tissues, transgenically in Transgenic hVim could be recovered from whole-fly lysates by using a standard procedure for intermediate filament (IF) extraction. When this procedure was used to test for the possible presence of IF-like proteins in flies, only lamins and tropomyosin were observed in IF-enriched extracts, thereby providing biochemical reinforcement to the paradigm that arthropods lack cIFs. In , transgenic hVim was unable to form filament networks in S2 cells and mesenchymal tissues; however, cage-like vimentin structures could be observed around the nuclei in internal epithelia, which suggests that retains selective competence for filament formation. Taken together, our results imply that although the filament network formation competence is partially lost in , a rudimentary filament network formation ability remains in epithelial cells. As a result of the observed selective competence for cIF assembly in , we hypothesize that internal epithelial cIFs were the last cIFs to disappear from arthropods.-Gullmets, J., Torvaldson, E., Lindqvist, J., Imanishi, S. Y., Taimen, P., Meinander, A., Eriksson, J. E. Internal epithelia in display rudimentary competence to form cytoplasmic networks of transgenic human vimentin.
细胞质中间丝(cIFs)存在于除节肢动物外的所有真后生动物中。为了研究节肢动物中cIFs的兼容性,我们在转基因果蝇中表达了人类波形蛋白(hVim),它是一种在脊椎动物细胞和组织中具有丝形成能力的cIF。通过使用中间丝(IF)提取的标准程序,可以从全蝇裂解物中回收转基因hVim。当使用该程序测试果蝇中可能存在的类IF蛋白时,在富含IF的提取物中仅观察到核纤层蛋白和原肌球蛋白,从而为节肢动物缺乏cIFs这一范式提供了生化证据。此外,转基因hVim在S2细胞和间充质组织中无法形成丝状网络;然而,在内皮中可以观察到细胞核周围有笼状波形蛋白结构,这表明其保留了丝状形成的选择性能力。综上所述,我们的结果表明,尽管在果蝇中丝状网络形成能力部分丧失,但上皮细胞中仍保留了基本的丝状网络形成能力。由于在果蝇中观察到了对cIF组装的选择性能力,我们推测内部上皮cIFs是节肢动物中最后消失的cIFs。-古尔梅茨,J.,托瓦尔森,E.,林德奎斯特,J.,今西,S.Y.,泰门,P.,梅南德,A.,埃里克松,J.E.果蝇的内部上皮显示出形成转基因人类波形蛋白细胞质网络的基本能力。