Skip Navigation



Plant and Cell Physiology Advance Access published online on February 23, 2008
Plant and Cell Physiology, doi:10.1093/pcp/pcn019
This Article
Right arrow Advance Access manuscript (PDF)
Right arrow All Versions of this Article:
49/3/420    most recent
pcn019v1
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Email this article to a friend
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Add to My Personal Archive
Right arrow Download to citation manager
Right arrowRequest Permissions
Google Scholar
Right arrow Articles by Abe, M.
Right arrow Articles by Shimamoto, K.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Abe, M.
Right arrow Articles by Shimamoto, K.
Agricola
Right arrow Articles by Abe, M.
Right arrow Articles by Shimamoto, K.
Social Bookmarking
Add to CiteULike   Add to Connotea   Add to Del.icio.us  
What's this?

© The Author 2008. 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

Identification of dynamin as an interactor of rice GIGANTEA by tandem-affinity purification (TAP)

Makoto Abe1, Masayuki Fujiwara1,2, Ken-ichi Kurotani1,3, Shuji Yokoi1,4 and Ko Shimamoto1

1Laboratory of Plant Molecular Genetics, Nara Institute of Science and Technology, 8916-5 Takayama, Ikoma 630-0101, Japan

Corresponding author: Ko Shimamoto, Laboratory of Plant Molecular Genetics, Nara Institute of Science and Technology, 8916-5 Takayama, Ikoma 630-0101, Japan. Phone: 0743 72 5500; Fax: 0743 72 5502; E-mail: simamoto{at}bs.naist.jp


  Abstract

GI, CO and FT regulate photoperiodic flowering in Arabidopsis. In rice, OsGI, Hd1 and Hd3a were identified as orthologs of GI, CO and FT, respectively, and are also important regulators of flowering. Although GI has roles in both flowering and the circadian clock, our understanding of its biochemical functions is still limited. In this study we purified novel OsGI interacting proteins by using the tandem affinity purification (TAP) method. The TAP method has been used effectively in a number of model species to isolate proteins that interact with proteins of interest. However, in plants, the TAP method has been used in only a few studies, and no novel proteins have previously been isolated by this method. We generated transgenic rice plants and cell cultures expressing a TAP-tagged version of OsGI. After a two-step purification procedure the interacting proteins were analyzed by mass spectrometry. Seven proteins, including dynamin, were identified as OsGI-interacting proteins. The interaction of OsGI with dynamin was verified by co-immunoprecipitation using a myc-tagged version of OsGI. Moreover, an analysis of Arabidopsis dynamin mutants indicated that although the flowering times of the mutants were not different from those of wild-type plants, an aerial rosette phenotype was observed in the mutants. We also found that OsGI is present in both the nucleus and the cytosol by western blot analysis and by transient assays. These results indicate that the TAP method is effective for the isolation of novel proteins that interact with target proteins in plants.

Keywords: Flowering - GI - tandem affinity purification (TAP) - proteomics - rice

2Present address: Plant Science Education Unit, Laboratory of Plant Protein Analysis, Nara Institute of Science and Technology, 8916-5 Takayama, Ikoma 630-0101, Japan

3Present address: Division of Integrated Life Science, Graduate School of Biostudies, Kyoto University, Sakyo-ku, Kyoto

4Present address: Faculty of Agriculture, Iwate University, Morioka 020-8550, Japan

(Received December 10, 2007; Accepted February 3, 2008)
Add to CiteULike CiteULike   Add to Connotea Connotea   Add to Del.icio.us Del.icio.us    What's this?




Disclaimer:
Please note that abstracts for content published before 1996 were created through digital scanning and may therefore not exactly replicate the text of the original print issues. All efforts have been made to ensure accuracy, but the Publisher will not be held responsible for any remaining inaccuracies. If you require any further clarification, please contact our Customer Services Department.