期刊
PLANT JOURNAL
卷 57, 期 4, 页码 680-689出版社
WILEY
DOI: 10.1111/j.1365-313X.2008.03721.x
关键词
phytochrome; nucleus; transport; cell free system; Acetabularia
资金
- Deutsche Forschungsgemeinschaft
- Human Frontiers Science Program [LT00631/2003-C]
- Hungarian Scientific Research Fund (OTKA) [60106]
- Howard Hughes Medical Institute (HHMI) International Research Fellowship
- Freiburg Institute for Advanced Studies (FRIAS) External Fellowship
Translocation from the cytosol to the nucleus is an essential step in phytochrome (phy) signal transduction. In the case of phytochrome A (phyA), this step occurs with the help of FHY1 (far-red-elongated hypocotyl 1), a specific transport protein. To investigate the components involved in phyA transport, we used a cell-free system that facilitates the controlled addition of transport factors. For this purpose, we isolated nuclei from the unicellular green algae Acetabularia acetabulum. These nuclei are up to 100 mu m in diameter and allow easy detection of imported proteins. Experiments with isolated nuclei of Acetabularia showed that FHY1 is sufficient for phyA transport. The reconstituted system demonstrates all the characteristics of phytochrome transport in Arabidopsis thaliana. In addition, FHY1 was also actively exported from the nucleus, consistent with its role as a shuttle protein in plants. Therefore, we believe that isolated Acetabularia nuclei may be used as a general tool to study nuclear transport of plant proteins.
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