Plant and Cell Physiology, 2004, Vol. 45, No. 4 416-426
© 2004 Oxford University Press
Function Analysis of Phototropin2 using Fern Mutants Deficient in Blue Light-Induced Chloroplast Avoidance Movement
1 SORST, Japan Science and Technology Corporation, 1-8, Honcho 4-chome, Kawaguchi, 332-0012 Japan
2 PRESTO, Japan Science and Technology Corporation, 1-8, Honcho 4-chome, Kawaguchi, 332-0012 Japan
3 Division of Biological Regulation and Photobiology, National Institute for Basic Biology, Okazaki, 444-8585 Japan
4 RIKEN Institute, Hirosawa 2-1, Wako, Saitama, Japan
5 Department of Biological Sciences, Graduate School of Science, Tokyo Metropolitan University, Hachioji, 192-0397 Japan
Gametophytes of the fern Adiantum capillus-veneris L. were mutagenized by heavy ion beam irradiation and screened for mutants lacking chloroplast avoidance movement under high intensity blue light. Mutants recovered include several with small deletions in the AcPHOT2 gene. The avoidance movement response in these mutants could be restored by transient expression of non-mutant AcPHOT2 cDNA, indicating that the chloroplast avoidance movement in this fern is mediated by the Acphot2 protein. Further functional analyses of the Acphot2 protein were performed using this transient assay for chloroplast avoidance movement. The results obtained suggest that the LOV2, but not the LOV1, domain of Acphot2 is essential for avoidance movement, and that several residues in the C-terminus of the kinase domain contribute to the avoidance response. The rate of dark reversion of the photo-activated LOV2 domain, which was calculated photometrically, was too fast to account for the lifetime of phot2 signal estimated from physiological responses. However, the rate of dark reversion of the combined domains of LOV1 and LOV2 did correspond to the lifetime of the signal, suggesting that LOV1 might have some function in this response, although it is not essential for playing a role as a photoreceptor.
6 Present address: Institute of Applied Biochemistry, University of Tsukuba, Tsukuba, 305-8572 Japan.
7 Corresponding author: E-mail, kagawa{at}agbi.tsukuba.ac.jp; Fax, +81-29-853-6270.
8 Present address: Gene Research Center, Tokyo University of Agriculture and Technology, Fuchu, 183-8509 Japan.
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