Skip to main content

Advertisement

SpringerLink
Log in

TMBP200, a XMAP215 homologue of tobacco BY-2 cells, has an essential role in plant mitosis

  • Original Article
  • Published:
Protoplasma Aims and scope Submit manuscript

Abstract

TMBP200 from tobacco BY-2 cells is a member of the highly conserved family of microtubule-associated proteins that includes Xenopus XMAP215, human TOGp, and Arabidopsis MOR1/GEM1. XMAP215 homologues have an essential role in spindle assembly and function in animals and yeast, but their role in plant mitosis is not fully clarified. Here, we show by immunoblot analysis that TMBP200 levels in synchronously cultured BY-2 cells increased when the cells entered mitosis, thus indicating that TMBP200 plays an important role in mitosis in tobacco. To investigate the role of TMBP200 in mitosis, we employed inducible RNA interference to silence TMBP200 expression in BY-2 cells. The resulting depletion of TMBP200 caused severe defects in bipolar spindle formation and resulted in the appearance of multinucleated cells with variable-sized nuclei. This finding indicates that TMBP200 has an essential role in bipolar spindle formation and function.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6

Similar content being viewed by others

References

  • Cassimeris L, Morabito J (2004) TOGp, the human homolog of XMAP215/Dis1, is required for centrosome integrity, spindle pole organization, and bipolar spindle assembly. Mol Biol Cell 215:1580–1590

    Article  Google Scholar 

  • Chan J, Jensen CG, Jensen LC, Bush M, Lloyd CW (1999) The 65-kDa carrot microtubule-associated protein forms regularly arranged filamentous cross-bridges between microtubules. Proc Natl Acad Sci USA 96:14931–14936

    Article  PubMed  CAS  Google Scholar 

  • Charrasse S, Schroeder M, Gauthier-Rouviere C, Ango F, Cassimeris L, Gard DL, Larroque C (1998) The TOGp protein is a new human microtubule-associated protein homologous to the Xenopus XMAP215. J Cell Sci 111:1371–1383

    PubMed  CAS  Google Scholar 

  • Conte N, Delaval B, Ginestier C, Ferrand A, Isnardon D, Larroque C, Prigent C, Séraphin B, Jacquemier J, Birnbaum D (2003) TACC1-chTOG-Aurora A protein complex in breast cancer. Oncogene 22:8102–8116

    Article  PubMed  CAS  Google Scholar 

  • Cullen CF, Deak P, Glover DM, Ohkura H (1999) Mini spindles: A gene encoding a conserved microtubule-associated protein required for the integrity of the mitotic spindle in Drosophila. J Cell Biol 146:1005–1018

    Article  PubMed  CAS  Google Scholar 

  • Curtis MD, Grossniklaus U (2003) A gateway cloning vector set for high-throughput functional analysis of genes in planta. Plant Physiol 133:462–469

    Article  PubMed  CAS  Google Scholar 

  • Eleftheriou EP, Baskin TI, Hepler PK (2005) Aberrant cell plate formation in the Arabidopsis thaliana microtubule organization 1 mutant. Plant Cell Physiol 46:671–675

    Article  PubMed  CAS  Google Scholar 

  • Gard DL, Becker BE, Josh Romney S (2004) MAPping the eukaryotic tree of life: structure, function, and evolution of the MAP215/Dis1 family of microtubule-associated proteins. Int Rev Cytol 239:179–272

    Article  PubMed  CAS  Google Scholar 

  • Gergely F, Draviam VM, Raff JW (2003) The ch-TOG/XMAP215 protein is essential for spindle pole organization in human somatic cells. Genes Dev 17:336–341

    Article  PubMed  CAS  Google Scholar 

  • Hamada T, Igarashi H, Itoh TJ, Shimmen T, Sonobe S (2004) Characterization of a 200 kDa microtubule-associated protein of tobacco BY-2 cells, a member of the XMAP215/MOR1 family. Plant Cell Physiol 45:1233–1242

    Article  PubMed  CAS  Google Scholar 

  • Hamada T, Itoh TJ, Hashimoto T, Shimmen T, Sonobe S (2009) GTP is required for the microtubule catastrophe-inducing activity of MAP200, a tobacco homolog of XMAP215. Plant Physiol 151:1823–1830

    Article  PubMed  CAS  Google Scholar 

  • Holmfeldt P, Stenmark S, Gullberg M (2004) Differential functional interplay of TOGp/XMAP215 and the KinI kinesin MCAK during interphase and mitosis. EMBO J 23:627–637

    Article  PubMed  CAS  Google Scholar 

  • Hush JM, Wadsworth P, Callaham DA, Hepler PK (1994) Quantification of microtubule dynamics in living plant cells using fluorescence redistribution after photobleaching. J Cell Sci 107:775–784

    PubMed  Google Scholar 

  • Jiang CJ, Sonobe S (1993) Identification and preliminary characterization of a 65 kDa higher-plant microtubule-associated protein. J Cell Sci 105:891–901

    CAS  Google Scholar 

  • Kakimoto T, Shibaoka H (1988) Cytoskeletal ultrastructure of phragmoplast–nuclei complexes isolated from cultured tobacco cells. Protoplasma Suppl 2:95–103

    Google Scholar 

  • Kawamura E, Wasteneys GO (2008) MOR1, the Arabidopsis thaliana homologue of Xenopus MAP215, promotes rapid growth and shrinkage, and suppresses the pausing of microtubules in vivo. J Cell Sci 121:4114–4123

    Article  PubMed  CAS  Google Scholar 

  • Kawamura E, Himmelspach R, Rashbrooke MC, Whittington AT, Gale KR, Collings DA, Wasteneys GO (2006) MICROTUBULE ORGANIZATION 1 regulates structure and function of microtubule arrays during mitosis and cytokinesis in the Arabidopsis root. Plant Physiol 140:102–114

    Article  PubMed  CAS  Google Scholar 

  • Kumagai F, Yoneda A, Tomida T, Sano T, Nagata T, Hasezawa S (2001) Fate of nascent microtubules organized at the M/G1 interface, as visualized by synchronized tobacco BY-2 cells stably expressing GFP-tubulin: time-sequence observations of the reorganization of cortical microtubules in living plant cells. Plant Cell Physiol 42:723–732

    Article  PubMed  CAS  Google Scholar 

  • Kurihara D, Matsunaga S, Uchiyama S, Fukui K (2008) Live cell imaging reveals plant aurora kinase has dual roles during mitosis. Plant Cell Physiol 49:1256–1261

    Article  PubMed  CAS  Google Scholar 

  • Niwa Y, Hirano T, Yoshimoto K, Shimizu M, Kobayashi H (1999) Non-invasive quantitative detection and applications of non-toxic, S65T-type green fluorescent protein in living plants. Plant J 18:455–463

    Article  PubMed  CAS  Google Scholar 

  • Shirasu-Hiza M, Coughlin P, Mitchison T (2003) Identification of XMAP215 as a microtubule-destabilizing factor in Xenopus egg extract by biochemical purification. J Cell Biol 161:349–358

    Article  PubMed  CAS  Google Scholar 

  • Tournebize R, Popov A, Kinoshita K, Ashford AJ, Rybina S, Pozniakovsky A, Mayer TU, Walczak CE, Karsenti E, Hyman AA (2000) Control of microtubule dynamics by the antagonistic activities of XMAP215 and XKCM1 in Xenopus egg extracts. Nat Cell Biol 2:13–19

    Article  PubMed  CAS  Google Scholar 

  • Twell D, Park SK, Hawkins TJ, Schubert D, Schmidt R, Smertenko A, Hussey PJ (2002) MOR1/GEM1 has an essential role in the plant-specific cytokinetic phragmoplast. Nat Cell Biol 4:711–714

    Article  PubMed  CAS  Google Scholar 

  • Weber K, Rathke PC, Osborn M (1978) Cytoplasmic microtubular images in glutaraldehyde-fixed tissue culture cells by electron microscopy and by immunofluorescence microscopy. Proc Natl Acad Sci USA 75:1820–1824

    Article  PubMed  CAS  Google Scholar 

  • Whittington AT, Vugrek O, Wei KJ, Hasenbein NG, Sugimoto K, Rashbrooke MC, Wasteneys GO (2001) MOR1 is essential for organizing cortical microtubules in plants. Nature 411:610–613

    Article  PubMed  CAS  Google Scholar 

  • Yasuhara H, Shibaoka H (2000) Inhibition of cell-plate formation by brefeldin A inhibited the depolymerization of microtubules in the central region of the phragmoplast. Plant Cell Physiol 41:300–310

    PubMed  CAS  Google Scholar 

  • Yasuhara H, Muraoka M, Shogaki H, Mori H, Sonobe S (2002) TMBP200, a microtubule bundling polypeptide isolated from telophase tobacco BY-2 cells is a MOR1 homologue. Plant Cell Physiol 43:595–603

    Article  PubMed  CAS  Google Scholar 

  • Yoneda A, Hasezawa S (2003) Origin of cortical microtubules organized at M/G1 interface: recruitment of tubulin from phragmoplast to nascent microtubules. Eur J Cell Biol 82:461–471

    Article  PubMed  CAS  Google Scholar 

Download references

Acknowledgements

We thank Dr. M. Curtis (University of Zürich, Switzerland) for providing pMDC7 and Dr. Y. Niwa (Shizuoka University, Japan) for the 35S-sGFP(S65T) vector. This work was supported by grant-in-aid for Scientific Research from the Ministry of Education, Culture, Sports, Science and Technology (no. 15770033 to H.Y.).

Conflicts of interest statement

The authors declare that they have no conflict of interest.

Author information

Author notes

  1. Yuki Oe

    Present address: Department of Neuroscience, Graduate School of Frontier Biosciences, Osaka University, Suita, Osaka, 565-0871, Japan

Authors and Affiliations

  1. Department of Life Science and Biotechnology, Faculty of Chemistry, Materials and Bioengineering, Kansai University, 3-3-35 Yamate-cho, Suita, Osaka, 564-8680, Japan

    Hiroki Yasuhara & Yuki Oe

Authors
  1. Hiroki Yasuhara
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Yuki Oe
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Hiroki Yasuhara.

Additional information

Handling Editor: Anne-Catherine Schmit

Electronic supplementary material

Below is the link to the electronic supplementary material.

709_2010_189_MOESM1_ESM.mov

Supplemental Movie 1 Time-lapse observation of the dynamics of microtubule localization during mitosis and cytokinesis in a living YTUB-TMRNAi-1 cell (an uninduced control cell) Images were recorded every 1 min. An image showing nuclear envelope breakdown set at 0 min. The progression of time before or after the breakdown is shown in the upper right corner of the images. (MOV 3036 kb)

Supplemental Movie 2 Time-lapse observation of the dynamics of microtubule localization during the inhibition of bipolar spindle formation in a living YTUB-TMRNAi-1 cell (an RNAi induced cell) Images were recorded every 1 min. An image showing nuclear envelope breakdown is set at 0 min. The progression of time before or after the breakdown is shown in the upper right corner of the images. (MOV 7177 kb)

Fig7

Supplemental Figure 1 Induction of RNAi silencing of YFP-fused β-tubulin did not affect the levels of TMBP200 and mitosis. YTB-GFPRNAi-1 cells were cultured for 3 days and then further cultured in the absence or presence of 5 μM β-estradiol to induce RNAi expression. At the indicated times after induction of RNAi expression, cells were harvested for protein extraction and also for staining with DAPI. Top Western blot with PAb200C, with anti-GFP antibodies, which also crossreact with YFP, and with ant-α-tubulin antibody as a loading control. Bottom Percentages of cells with a single nucleus (1), mitotic cells (mitotic), binucleated cells (2), and multinucleated cells (multi.). (GIF 24 kb)

High resolution image (TIFF 14416 kb)

Rights and permissions

Reprints and permissions

About this article

Cite this article

Yasuhara, H., Oe, Y. TMBP200, a XMAP215 homologue of tobacco BY-2 cells, has an essential role in plant mitosis. Protoplasma 248, 493–502 (2011). https://doi.org/10.1007/s00709-010-0189-6

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00709-010-0189-6

Keywords

Search

Navigation