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Published on April 22, 2009; 10.1104/pp.108.130757

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Received October 3, 2008
Accepted April 6, 2009

Cytochrome P450s as reporters for circadian-regulated pathways

Yinghong Pan , Todd P. Michael , Matthew E. Hudson , Steve A. Kay , Joanne Chory , and Mary A. Schuler *

Department of Cell and Developmental Biology, University of Illinois, Urbana, IL 61801; Department of Plant Biology and Pathology, Waksman Institute for Microbiology, Rutgers University, Piscataway, NJ 08854; Department of Crop Sciences, University of Illinois, Urbana, IL 61801; Section of Cell and Developmental Biology, University of California San Diego, La Jolla CA 92093; Plant Biology Laboratory and Howard Hughes Medical Institute, Salk Institute for Biological Studies, La Jolla, CA 92037

* Corresponding author; email: maryschu{at}uiuc.edu.

Cytochrome P450 monooxygenases play important roles in the synthesis of diverse secondary compounds in Arabidopsis thaliana. Comparison of four datasets analyzing seedlings harvested over a two-day period of constant conditions after growth with varying photoperiods and thermocycles recorded a total of 98 P450 loci as circadian-regulated for at least one of the four conditions. Here, we further describe the circadian-regulated pathways using, as reporters, individual P450 loci that are likely to be rate-limiting in secondary metabolic pathways. RT-PCR gel blot analyses have confirmed circadian regulation of P450s in phenylpropanoid, carotenoid, oxylipin, glucosinolate and brassinosteroid biosyntheses, and shown that both P450 and non-P450 genes in the many branches of the phenylpropanoid pathway have similar circadian patterns of expression. In silico analyses of the subsets of co-regulated promoters have identified over-represented promoter elements in various biosynthetic pathway genes including MYB and MYB4 elements that are significantly more abundant in promoters for the core and lignin branch of phenylpropanoid metabolism. Interactions with these elements important for circadian regulation do not involve the MYB transcription factor PAP1 as previously proposed since the expression patterns of circadian-regulated P450s are the same in pap1-D mutant seedlings as in wildtype seedlings. Further analysis of circadian-regulated promoters in other biochemical pathways provides us with the opportunity to identify novel promoter motifs that might be important in P450 circadian regulation.






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