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© The Rockefeller University Press, 0021-9525/1999//437 $5.00
The Journal of Cell Biology, Volume 145, Number 3, , 1999 437-445

Light-dependent Translocation of a Phytochrome B-GFP Fusion Protein to the Nucleus in Transgenic Arabidopsis

Rumi Yamaguchi^*, Masanobu Nakamura^*, Nobuyoshi Mochizuki^*, Steve A. Kay, and Akira Nagatani^*

^* Department of Botany, Graduate School of Science, Kyoto University, Kyoto 606-8502, Japan; and National Science Foundation Center for Biological Timing, Department of Cell Biology, Scripps Research Institute, La Jolla, California 92037

Phytochrome is a ubiquitous photoreceptor of plants and is encoded by a small multigene family. We have shown recently that a functional nuclear localization signal may reside within the COOH-terminal region of a major member of the family, phytochrome B (phyB) (Sakamoto, K., and A. Nagatani. 1996. Plant J. 10:859–868). In the present study, a fusion protein consisting of full-length phyB and the green fluorescent protein (GFP) was overexpressed in the phyB mutant of Arabidopsis to examine subcellular localization of phyB in intact tissues. The resulting transgenic lines exhibited pleiotropic phenotypes reported previously for phyB overexpressing plants, suggesting that the fusion protein is biologically active. Immunoblot analysis with anti-phyB and anti-GFP monoclonal antibodies confirmed that the fusion protein accumulated to high levels in these lines. Fluorescence microscopy of the seedlings revealed that the phyB-GFP fusion protein was localized to the nucleus in light grown tissues. Interestingly, the fusion protein formed speckles in the nucleus. Analysis of confocal optical sections confirmed that the speckles were distributed within the nucleus. In contrast, phyB-GFP fluorescence was observed throughout the cell in dark-grown seedlings. Therefore, phyB translocates to specific sites within the nucleus upon photoreceptor activation.

Key Words: green fluorescent protein • nuclear targeting • photomorphogenesis • phytochrome • signal transduction

Abbreviations used in this paper: DIC, differential interference contrast; GFP, green fluorescent protein; Pfr, far-red light absorbing form of phytochrome; phy, phytochrome.

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• Zhu, Y., Tepperman, J. M., Fairchild, C. D., Quail, P. H. (2000). Phytochrome B binds with greater apparent affinity than phytochrome A to the basic helix-loop-helix factor PIF3 in a reaction requiring the PAS domain of PIF3. Proc. Natl. Acad. Sci. USA 10.1073/pnas.230433797v1 [Abstract] [Full Text]  
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• Fairchild, C. D., Schumaker, M. A., Quail, P. H. (2000). HFR1 encodes an atypical bHLH protein that acts in phytochrome A signal transduction. Genes Dev. 14: 2377-2391 [Abstract] [Full Text]  
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• Cerdán, P. D., Staneloni, R. J., Ortega, J., Bunge, M. M., Rodriguez-Batiller, M. J., Sánchez, R. A., Casal, J. J. (2000). Sustained but Not Transient Phytochrome A Signaling Targets a Region of an Lhcb1*2 Promoter Not Necessary for Phytochrome B Action. Plant Cell 12: 1203-1212 [Abstract] [Full Text]  
• Bolle, C., Koncz, C., Chua, N.-H. (2000). PAT1, a new member of the GRAS family, is involved in phytochrome A signal transduction. Genes Dev. 14: 1269-1278 [Abstract] [Full Text]  
• Martínez-García, J. F., Huq, E., Quail, P. H. (2000). Direct Targeting of Light Signals to a Promoter Element-Bound Transcription Factor. Science 288: 859-863 [Abstract] [Full Text]  
• Reed, J. W., Nagpal, P., Bastow, R. M., Solomon, K. S., Dowson-Day, M. J., Elumalai, R. P., Millar, A. J. (2000). Independent Action of ELF3 and phyB to Control Hypocotyl Elongation and Flowering Time. Plant Physiol. 122: 1149-1160 [Abstract] [Full Text]  
• Neff, M. M., Fankhauser, C., Chory, J. (2000). Light: an indicator of time and place. Genes Dev. 14: 257-271 [Full Text]  
• Bognar, L. K., Hall, A., Adam, E., Thain, S. C., Nagy, F., Millar, A. J. (1999). The circadian clock controls the expression pattern of the circadian input photoreceptor, phytochrome B. Proc. Natl. Acad. Sci. USA 96: 14652-14657 [Abstract] [Full Text]  
• Ku, N.-O., Zhou, X., Toivola, D. M., Omary, M. B. (1999). The cytoskeleton of digestive epithelia in health and disease. Am. J. Physiol. Gastrointest. Liver Physiol. 277: G1108-G1137 [Abstract] [Full Text]  
• Parks, B. M., Spalding, E. P. (1999). Sequential and coordinated action of phytochromes A and B during Arabidopsis stem growth revealed by kinetic analysis. Proc. Natl. Acad. Sci. USA 96: 14142-14146 [Abstract] [Full Text]  
• Stacey, M. G., von Arnim, A. G. (1999). A Novel Motif Mediates the Targeting of the Arabidopsis COP1 Protein to Subnuclear Foci. J. Biol. Chem. 274: 27231-27236 [Abstract] [Full Text]  
• Zhu, Y., Tepperman, J. M., Fairchild, C. D., Quail, P. H. (2000). Phytochrome B binds with greater apparent affinity than phytochrome A to the basic helix-loop-helix factor PIF3 in a reaction requiring the PAS domain of PIF3. Proc. Natl. Acad. Sci. USA 97: 13419-13424 [Abstract] [Full Text]  

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