Journal of Plant Research

, Volume 129, Issue 6, pp 1165–1178

Plant Aurora kinases interact with and phosphorylate transcription factors

  • Mai Takagi
  • Takuya Sakamoto
  • Ritsuko Suzuki
  • Keiichirou Nemoto
  • Takeshi Obayashi
  • Takeshi Hirakawa
  • Tomoko M. Matsunaga
  • Daisuke Kurihara
  • Yuko Nariai
  • Takeshi Urano
  • Tatsuya Sawasaki
  • Sachihiro Matsunaga
Regular Paper
  • 464 Downloads

Abstract

Aurora kinase (AUR) is a well-known mitotic serine/threonine kinase that regulates centromere formation, chromosome segregation, and cytokinesis in eukaryotes. In addition to regulating mitotic events, AUR has been shown to regulate protein dynamics during interphase in animal cells. In contrast, there has been no identification and characterization of substrates and/or interacting proteins during interphase in plants. The Arabidopsis thaliana genome encodes three AUR paralogues, AtAUR1, AtAUR2, and AtAUR3. Among them, AtAUR1 and AtAUR2 are considered to function redundantly. Here, we confirmed that both AtAUR1 and AtAUR3 are localized in the nucleus and cytoplasm during interphase, suggesting that they have functions during interphase. To identify novel interacting proteins, we used AlphaScreen to target 580 transcription factors (TFs) that are mainly functional during interphase, using recombinant A. thaliana TFs and AtAUR1 or AtAUR3. We found 133 and 32 TFs had high potential for interaction with AtAUR1 and AtAUR3, respectively. The highly AtAUR-interacting TFs were involved in various biological processes, suggesting the functions of the AtAURs during interphase. We found that AtAUR1 and AtAUR3 showed similar interaction affinity to almost all TFs. However, in some cases, the interaction affinity differed substantially between the two AtAUR homologues. These results suggest that AtAUR1 and AtAUR3 have both redundant and distinct functions through interactions with TFs. In addition, database analysis revealed that most of the highly AtAUR-interacting TFs contained a detectable phosphopeptide that was consistent with the consensus motifs for human AURs, suggesting that these TFs are substrates of the AtAURs. The AtAURs phosphorylated several highly interacting TFs in the AlphaScreen in vitro. Overall, in line with the regulation of TFs through interaction, our results indicate the possibility of phosphoregulation of several TFs by the AtAURs (280/300).

Keywords

Aurora kinase Transcription factors AlphaScreen Wheat germ cell-free protein synthesis In vitro phosphorylation assay with ATPγS 

Supplementary material

10265_2016_860_MOESM1_ESM.pdf (1.6 mb)
Supplementary material 1 (PDF 1665 kb)

Copyright information

© The Botanical Society of Japan and Springer Japan 2016

Authors and Affiliations

  • Mai Takagi
    • 1
  • Takuya Sakamoto
    • 1
  • Ritsuko Suzuki
    • 1
  • Keiichirou Nemoto
    • 2
  • Takeshi Obayashi
    • 3
  • Takeshi Hirakawa
    • 1
  • Tomoko M. Matsunaga
    • 4
  • Daisuke Kurihara
    • 5
  • Yuko Nariai
    • 6
  • Takeshi Urano
    • 6
  • Tatsuya Sawasaki
    • 2
  • Sachihiro Matsunaga
    • 1
  1. 1.Department of Applied Biological Science, Faculty of Science and TechnologyTokyo University of ScienceNodaJapan
  2. 2.Proteo-Science CenterEhime UniversityMatsuyamaJapan
  3. 3.Graduate School of Information SciencesTohoku UniversitySendaiJapan
  4. 4.Institute for Science and TechnologyTokyo University of ScienceNodaJapan
  5. 5.Division of Biological Science, Graduate School of ScienceNagoya University, JST ERATO Higashiyama Live-Holonics ProjectNagoyaJapan
  6. 6.Department of Biochemistry, Faculty of MedicineShimane UniversityIzumoJapan

Personalised recommendations