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

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

3 Scopus citations

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 (US)
Pages (from-to)431-439
Number of pages9
JournalJournal of Biochemistry
Volume153
Issue number5
DOIs
StatePublished - May 2013

Keywords

  • 14-3-3
  • Trypanosoma brucei
  • cell division
  • cytokinesis
  • dimerization

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology

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