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Published on September 6, 2002; 10.1104/pp.004929

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Received February 26, 2002
Returned for revision May 22, 2002
Accepted June 10, 2002

Phase-Specific Circadian Clock Regulatory Elements in Arabidopsis

Todd P. Michael and C. Robertson McClung *

Department of Biological Sciences, Dartmouth College, Hanover, New Hampshire 03755

* Corresponding author; email: mcclung{at}dartmouth.edu.

We have defined a minimal Arabidopsis CATALASE 3 (CAT3) promoter sufficient to drive evening-specific circadian transcription of a LUCIFERASE reporter gene. Deletion analysis and site-directed mutagenesis reveal a circadian response element, the evening element (EE: AAAATATCT), that is necessary for evening-specific transcription. The EE differs only by a single base pair from the CIRCADIAN CLOCK ASSOCIATED 1-binding site (CBS: AAAAAATCT), which is important for morning-specific transcription. We tested the hypothesis that the EE and the CBS specify circadian phase by site-directed mutagenesis to convert the CAT3 EE into a CBS. Changing the CAT3 EE to a CBS changes the phase of peak transcription from the evening to the morning in continuous dark and in light-dark cycles, consistent with the specification of phase by the single base pair that distinguishes these elements. However, rhythmicity of the CBS-containing CAT3 promoter is dramatically compromised in continuous light. Thus, we conclude that additional information normally provided in the context of a morning-specific promoter is necessary for full circadian activity of the CBS.




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