Abstract
The circadian clock is required for adaptive responses to daily and seasonal changes in environmental conditions1,2,3. Light and the circadian clock interact to consolidate the phase of hypocotyl cell elongation to peak at dawn under diurnal cycles in Arabidopsis thaliana4,5,6,7. Here we identify a protein complex (called the evening complex)—composed of the proteins encoded by EARLY FLOWERING 3 (ELF3), ELF4 and the transcription-factor-encoding gene LUX ARRHYTHMO (LUX; also known as PHYTOCLOCK 1)—that directly regulates plant growth8,9,10,11,12. ELF3 is both necessary and sufficient to form a complex between ELF4 and LUX, and the complex is diurnally regulated, peaking at dusk. ELF3, ELF4 and LUX are required for the proper expression of the growth-promoting transcription factors encoded by PHYTOCHROME INTERACTING FACTOR 4 (PIF4) and PIF5 (also known as PHYTOCHROME INTERACTING FACTOR 3-LIKE 6) under diurnal conditions4,6,13. LUX targets the complex to the promoters of PIF4 and PIF5 in vivo. Mutations in PIF4 and/or PIF5 are epistatic to the loss of the ELF4–ELF3–LUX complex, suggesting that regulation of PIF4 and PIF5 is a crucial function of the complex. Therefore, the evening complex underlies the molecular basis for circadian gating of hypocotyl growth in the early evening.
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Acknowledgements
P. Y. Pongsawakul illustrated the model in Fig. 4c. We thank G. Breton, T. Hirota, J. Pruneda-Paz, D. Nagel, E. Kolmos and B. J. Cole for critical reading of the manuscript. We also thank J. Halverson, A. L. Quiroz and C. Valdivia for excellent technical assistance. pif4-101 pif5-1 seedlings were a generous gift from S. Lorrain and C. Fankhauser. F. Harmon originally identified the nature of the elf4-3 mutation and designed the dCAPS strategy. This work was supported by a University of California, San Diego, Chancellor’s Undergraduate Research Scholarship (to J.J.K.), grants from the European Molecular Biology Organization (ALTF 236-2005 to A.H.), the National Science Foundation (IBN-0416762 to T.F.S.) and the National Institutes of Health (NRSA GM083585 to D.A.N., NRSA GM080930 to E.E.H., R01 GM79712 to T.I., and R01 GM50006 and GM67837 to S.A.K.).
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D.A.N., A.H., E.E.H., T.I., T.F.S., E.M.F. and S.A.K. designed the experiments. D.A.N. performed and analysed all of the immunoprecipitations and ChIP assays, generated and characterized the anti-ELF3 antibody and the transgenic lines, and generated the plasmids for yeast three-hybrid analysis. D.A.N. also co-performed the gene expression analysis with A.H. A.H. performed the yeast two- and one-hybrid assays and generated and characterized the LUX and LUX/NOX ami transgenic lines. E.E.H. generated the anti-LUX antibody, characterized transgenic lines and co-performed western blot analysis with D.A.N. J.J.K. measured hypocotyls, performed and analysed the yeast three-hybrid assay, and assisted with the yeast one-hybrid analysis and generation of the elf3-2 pif4-101 pif5-1 seedlings. T.I. performed the original yeast two-hybrid assays. T.F.S. generated the ELF4::ELF4–HA elf4-2 line. E.M.F. characterized the ELF4-transgenic lines. D.A.N. and S.A.K. wrote the manuscript.
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Nusinow, D., Helfer, A., Hamilton, E. et al. The ELF4–ELF3–LUX complex links the circadian clock to diurnal control of hypocotyl growth. Nature 475, 398–402 (2011). https://doi.org/10.1038/nature10182
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DOI: https://doi.org/10.1038/nature10182
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