Trypanosoma brucei 14-3-3I and II proteins predominantly form a heterodimer structure that acts as a potent cell cycle regulator in vivo

Masahiro Inoue, Kouichi Yasuda, Haruki Uemura, Natsumi Yasaka, Achim Schnaufer, Mihiro Yano, Hiroshi Kido, Daisuke Kohda, Hirofumi Doi, Toshihide Fukuma, Akihiko Tsuji, Nobuo Horikoshi

    Research output: Contribution to journalArticle

    3 Citations (Scopus)

    Abstract

    Hetero- and homodimerization of 14-3-3 proteins demonstrate distinctive functions in mammals and plants. Trypanosoma brucei 14-3-3I and II (Tb14-3-3I and II) play pivotal roles in motility, cytokinesis and the cell cycle; however, the significance and the mechanism of Tb14-3-3 dimerization are remained to be elucidated. We found that ectopically expressed epitope-tagged Tb14-3-3I and II proteins formed hetero- and homodimers with endogenous Tb14-3-3I and II proteins. However, we also found the ability to form hetero- or homodimers between Tb14-3-3I and II proteins was clearly affected by the sequence and location of the epitope tag used. We found a blue native polyacrylamide gel electrophoresis system followed by western blotting may distinguish monomer from dimer structure, and stable from unstable conformation of Tb14-3-3. Combined with co-immunoprecipitation results, we revealed that Tb14-3-3 proteins mainly existed as heterodimeric form. Furthermore, co-overexpression of Tb14-3-3I and II proteins in T. brucei induced aberrant numbers of organelles in cells, but overexpression of either isoform alone rarely produced such morphology. These results suggest that heterodimers play more significant roles than homodimers not only in the maintenance of steady-state levels of the 14-3-3 proteins but also in the regulation of cytokinesis.

    Original languageEnglish
    Pages (from-to)431-439
    Number of pages9
    JournalJournal of biochemistry
    Volume153
    Issue number5
    DOIs
    Publication statusPublished - May 1 2013

    Fingerprint

    Trypanosoma brucei brucei
    Cell Cycle
    Cells
    14-3-3 Proteins
    Cytokinesis
    Proteins
    Epitopes
    Native Polyacrylamide Gel Electrophoresis
    Mammals
    Dimerization
    Electrophoresis
    Immunoprecipitation
    Organelles
    Dimers
    Conformations
    Protein Isoforms
    Monomers
    Western Blotting
    Maintenance

    All Science Journal Classification (ASJC) codes

    • Biochemistry
    • Molecular Biology

    Cite this

    Trypanosoma brucei 14-3-3I and II proteins predominantly form a heterodimer structure that acts as a potent cell cycle regulator in vivo. / Inoue, Masahiro; Yasuda, Kouichi; Uemura, Haruki; Yasaka, Natsumi; Schnaufer, Achim; Yano, Mihiro; Kido, Hiroshi; Kohda, Daisuke; Doi, Hirofumi; Fukuma, Toshihide; Tsuji, Akihiko; Horikoshi, Nobuo.

    In: Journal of biochemistry, Vol. 153, No. 5, 01.05.2013, p. 431-439.

    Research output: Contribution to journalArticle

    Inoue, M, Yasuda, K, Uemura, H, Yasaka, N, Schnaufer, A, Yano, M, Kido, H, Kohda, D, Doi, H, Fukuma, T, Tsuji, A & Horikoshi, N 2013, 'Trypanosoma brucei 14-3-3I and II proteins predominantly form a heterodimer structure that acts as a potent cell cycle regulator in vivo', Journal of biochemistry, vol. 153, no. 5, pp. 431-439. https://doi.org/10.1093/jb/mvt016
    Inoue, Masahiro ; Yasuda, Kouichi ; Uemura, Haruki ; Yasaka, Natsumi ; Schnaufer, Achim ; Yano, Mihiro ; Kido, Hiroshi ; Kohda, Daisuke ; Doi, Hirofumi ; Fukuma, Toshihide ; Tsuji, Akihiko ; Horikoshi, Nobuo. / Trypanosoma brucei 14-3-3I and II proteins predominantly form a heterodimer structure that acts as a potent cell cycle regulator in vivo. In: Journal of biochemistry. 2013 ; Vol. 153, No. 5. pp. 431-439.
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