Plant Molecular Biology

, Volume 74, Issue 6, pp 537–547

Tobacco microtubule-associated protein, MAP65-1c, bundles and stabilizes microtubules

  • Qiutao Meng
  • Jizhou Du
  • Jiejie Li
  • Xiaomei Lü
  • Xian Zeng
  • Ming Yuan
  • Tonglin Mao
Article

Abstract

Three genes that encode MAP65-1 family proteins have been identified in the Nicotiana tabacum genome. In this study, NtMAP65-1c fusion protein was shown to bind and bundle microtubules (MTs). Further in vitro investigations demonstrated that NtMAP65-1c not only alters MT assembly and nucleation, but also exhibits high MT stabilizing activity against cold or katanin-induced destabilization. Analysis of NtMAP65-1c-GFP expressing BY-2 cells clearly demonstrated that NtMAP65-1c was able to bind to MTs during specific stages of the cell cycle. Furthermore, in vivo, NtMAP65-1c-GFP-bound cortical MTs displayed an increase in resistance against the MT-disrupting drug, propyzamide, as well as against cold temperatures. Taken together, these results strongly suggest that NtMAP65-1c stabilizes MTs and is involved in the regulation of MT organization and cellular dynamics.

Keywords

NtMAP65-1c Microtubules-associated protein Microtubule stabilization Katanin 

Abbreviations

MAPs

Microtubule-associated proteins

PIPES

Piperazine-N,N′-bis (2-ethanesulfonic acid 1,4-piperazinediethanesulfonic acid)

EGTA

Ethylene glycol-bis-(β-aminoethyl ether)-N,N,N′,N′-tetraacetic acid

Supplementary material

11103_2010_9694_MOESM1_ESM.jpg (575 kb)
NtMAP65-1c decreased the critical concentration (Cc) for tubulin polymerization. (a) Tubulins at concentration of 5 or 10 μM underwent polymerization in presence or absence of 2 μM NtMAP65-1c at 35°C for 30 min. After centrifuging at 100,000 g for 20 min, the supernatant and pellet were subjected to SDS–PAGE. Tubulins assembled without GTP were used a control. (b) The amount of MTs in the pellet of (a) was estimated following density scanning of gels. There was a notable increase in the amount of tubulin in the pellet in the presence of NtMAP65-1c, as compared to control. (JPEG 574 kb)
11103_2010_9694_MOESM2_ESM.jpg (586 kb)
NtMAP65-1a is capable of stabilizing MTs against in vitro cold treatment. Tubulins (20 μM) were assembled with NtMAP65-1a fusion protein at 35°C for 20 min and then incubated at 4°C for 20 min. The pellets were subjected to SDS–PAGE. As a positive control, paclitaxel (10 μM) was used in place of NtMAP65-1a. (JPEG 586 kb)

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Copyright information

© Springer Science+Business Media B.V. 2010

Authors and Affiliations

  • Qiutao Meng
    • 1
  • Jizhou Du
    • 1
  • Jiejie Li
    • 1
  • Xiaomei Lü
    • 1
  • Xian Zeng
    • 1
  • Ming Yuan
    • 1
  • Tonglin Mao
    • 1
  1. 1.State Key Laboratory of Plant Physiology and Biochemistry; Department of Plant SciencesCollege of Biological Sciences, China Agricultural UniversityBeijingChina

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