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First published online May 27, 2005; 10.1105/tpc.105.033035

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tpc.105.033035v1
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The Plant Cell 17:1926-1940 (2005)
© 2005 American Society of Plant Biologists

Positive and Negative Factors Confer Phase-Specific Circadian Regulation of Transcription in Arabidopsis{boxw}

Stacey L. Harmera,1 and Steve A. Kayb

a Section of Plant Biology, University of California, Davis, Davis, California, 95616
b Department of Cell Biology, Scripps Research Institute, La Jolla, California, 92037

1 To whom correspondence should be addressed. E-mail slharmer{at}ucdavis.edu; fax 530-752-5410.

The circadian clock exerts a major influence on transcriptional regulation in plants and other organisms. We have previously identified a motif called the evening element (EE) that is overrepresented in the promoters of evening-phased genes. Here, we demonstrate that multimerized EEs are necessary and sufficient to confer evening-phased circadian regulation. Although flanking sequences are not required for EE function, they can modulate EE activity. One flanking sequence, taken from the PSEUDORESPONSE REGULATOR 9 promoter, itself confers dawn-phased rhythms and has allowed us to define a new clock promoter motif (the morning element [ME]). Scanning mutagenesis reveals that both activators and repressors of gene expression act through the ME and EE. Although our experiments confirm that CIRCADIAN CLOCK ASSOCIATED 1 (CCA1) and LATE ELONGATED HYPOCOTYL (LHY) are likely to act as repressors via the EE, they also show that they have an unexpected positive effect on EE-mediated gene expression as well. We have identified a clock-regulated activity in plant extracts that binds specifically to the EE and has a phase consistent with it being an activator of expression through the EE. This activity is reduced in CCA1/LHY null plants, suggesting it may itself be part of a circadian feedback loop and perhaps explaining the reduction in EE activity in these double mutant plants.




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