First published online June 10, 2005; 10.1104/pp.104.057414
Plant Physiology 138:1644-1652 (2005)
© 2005 American Society of Plant Biologists
CELL BIOLOGY AND SIGNAL TRANSDUCTION
A Sphingolipid Elicitor-Inducible Mitogen-Activated Protein Kinase Is Regulated by the Small GTPase OsRac1 and Heterotrimeric G-Protein in Rice1,[w]
Damien Lieberherr2,
Nguyen Phuong Thao,
Ayako Nakashima,
Kenji Umemura,
Tsutomu Kawasaki and
Ko Shimamoto*
Laboratory of Plant Molecular Genetics, Nara Institute of Science and Technology, Ikoma, Nara 630–0101, Japan (D.L., N.P.T., A.N., T.K., K.S.); and Agricultural and Veterinary Research Laboratories, Meiji Seika Kaisha, Ltd., Kohoku-ku, Yokohama 222–8567, Japan (K.U.)
Mitogen-activated protein kinase (MAPK) cascades are activated in plants during responses to pathogens or to pathogen-derived elicitors and mediate intracellular stress responses. Here, we show that a rice (Oryza sativa) MAPK, OsMAPK6, was posttranslationally activated in a cell culture by a sphingolipid elicitor. Suppression of OsMAPK6 expression by RNA interference resulted in a strong reduction of pathogen-induced Phe ammonia-lyase mRNA, whereas the mRNA level of another rice MAPK, OsMAPK5a, was highly increased. Silencing of a small GTPase, OsRac1, by RNA interference or loss-of-function mutation (d1) of the heterotrimeric G-protein  -subunit gene resulted in a strong reduction of the OsMAPK6 protein levels and of kinase activation by a sphingolipid elicitor. Furthermore, coimmunoprecipitation experiments with OsRac1 and OsMAPK6 proteins showed that OsMAPK6 is closely associated with the active form of OsRac1, but not with inactive forms of OsRac1. These results indicate that these two G-proteins regulate an elicitor-inducible MAPK in rice at the protein level.
1 This work was supported by the Japanese Society for the Promotion of Science (JSPS; postdoctoral fellowship no. P01701 to D.L.), the Research for the Future Program of the JSPS (grant no. JSPS–RFTF 00L01604), and the Ministry of Agriculture, Forestry, and Fisheries of Japan, Rice Genome Project.
2 Present address: Swiss Institute of Bioinformatics, Centre Medical Universitaire, Rue Michel Servet 1, 1211 Geneva 4, Switzerland.
[w] The online version of this article contains Web-only data.
Article, publication date, and citation information can be found at www.plantphysiol.org/cgi/doi/10.1104/pp.104.057414.
* Corresponding author; e-mail simamoto{at}bs.naist.jp; fax 81–743–72–5509.
Received November 29, 2004;
returned for revision March 30, 2005;
accepted May 3, 2005.
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