First published online September 6, 2002; 10.1104/pp.004929
Plant Physiol, October 2002, Vol. 130, pp. 627-638
Phase-Specific Circadian Clock Regulatory Elements in
Arabidopsis1
Todd P.
Michael and
C. Robertson
McClung*
Department of Biological Sciences, Dartmouth College, Hanover, New
Hampshire 03755
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.
1
This work was supported by the National Science
Foundation (grant no. IBN 9817603 to C.R.M.), by the U.S. Department of
Agriculture (National Research Initiative-Competitive Grants Program
grant no. 9602632 to C.R.M.), and by an institutional grant from the American Cancer Society to the Norris Cotton Cancer Center at Dartmouth College.
*
Corresponding author; e-mail mcclung{at}dartmouth.edu; fax
603-646-1347.
© 2002 American Society of Plant Physiologists
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