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Plant and Cell Physiology Advance Access published online on February 13, 2009
Plant and Cell Physiology, doi:10.1093/pcp/pcp024
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© The Author 2009. Published by Oxford University Press on behalf of Japanese Society of Plant Physiologists. All rights reserved. For Permissions, please e-mail: journals.permissions@oxfordjournals.org

The putative RNA-processing protein, THO2, is a microtubule-associated protein in tobacco.

Takahiro Hamada1,2,4, Hisako Igarashi1,3,4, Ryoichi Taguchi1, Masayuki Fujiwara2, Yoichiro Fukao2, Teruo Shimmen1, Etsuo Yokota1 and Seiji Sonobe1

1Department of Life Science, Graduate School of Life Science, University of Hyogo, Harima Science Park City, Hyogo, 678-1297, Japan
2Graduate School of Biological Sciences, Nara Institute of Science and Technology, Nara, 630-0101, Japan
3National Institute for Basic Biology, Myodaiji-cho, Okazaki, Aichi 444-8585, Japan

Corresponding Author: Dr. Takahiro Hamada. Graduate School of Biological Sciences, Nara Institute of Science and Technology, Nara, 630-0101, Japan, Phone number: +81-743-72-5526, Fax number: +81-743-72-5529, E-mail address: hama{at}bs.naist.jp


  Abstract

THO2 is a component of the THO/TREX complex that participates in mRNA metabolism and export from the nucleus in yeast and animal cells. Here we report that tobacco putative THO2-related protein (NtTHO2) is a microtubule-associated protein, which directly binds to microtubules in vitro and colocalizes with cortical microtubules in vivo. We purified endogenous NtTHO2 by cycles of microtubule polymerization-depolymerization from crude extracts of tobacco BY-2 miniprotoplasts. Purified NtTHO2 sedimented with microtubules in vitro. Immunofluorescence revealed that NtTHO2 was present in both nucleus and cytoplasm. In interphase, cytoplasmic NtTHO2 was localized along cortical microtubules. In mitotic phase, NtTHO2 was localized to the mitotic spindle but to neither the preprophase band nor phragmoplast. In mature cells of seedling roots, and in BY-2 cells in which proliferation was stopped by removing 2,4-D. NtTHO2 staining was confined mainly to the nucleolus. These results suggest that NtTHO2 is a multifunctional protein that participates in mRNA metabolism, and also functions within the cortical microtubules and mitotic spindle.

Keywords: BY-2 cells - Microtubule - Microtubule-associated protein - Nicotiana tabacum - nucleolus - THO2

4These authors contributed equally to this work.

(Received December 16, 2008; Accepted February 10, 2009)
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