Plant Physiology Preview
Published on October 16, 2003; 10.1104/pp.103.025114
Received April 9, 2003
Returned for revision May 23, 2003
Accepted July 10, 2003
HY5, Circadian Clock-Associated 1, and a cis-Element, DET1 Dark Response Element, Mediate DET1 Regulation of Chlorophyll a/b-Binding Protein 2 Expression
Bridey B. Maxwell , Carol R. Andersson , Daniel S. Poole , Steve A. Kay , and Joanne Chory *
Plant Biology Laboratory, The Salk Institute for Biological Studies, La Jolla, California 92037 (B.B.M., D.S.P., J.C.); Department of Biology, University of California, San Diego, California 92093 (B.B.M., D.S.P.); Department of Cell Biology and Institute for Childhood and Neglected Diseases, The Scripps Research Institute, La Jolla, California 92037 (C.R.A., S.A.K.); and Howard Hughes Medical Institute, The Salk Institute, La Jolla, California 92037 (J.C.)
* Corresponding author; email: chory{at}salk.edu.
DET1 is a pleiotropic regulator of Arabidopsis development and controls the expression of many light-regulated genes. To gain a better understanding of the mechanism by which DET1 controls transcription from light-regulated promoters, we identified elements in the chlorophyll a/b-binding protein 2 (CAB2) promoter that are required for DET1-mediated expression. Using a series of reporter constructs in which the luciferase gene is controlled by CAB2 promoter fragments, we defined two DET1-responsive elements in the CAB2 promoter that are essential for proper CAB2 transcription. A 40-bp DET1 dark-response element (DtRE) is required for both dark and root-specific repression of CAB2, whereas the known CAB upstream factor-1 element is required for DET1 activation-associated effects in the light and repression in the roots. HY5, a factor that binds CAB upstream factor-1, is also required for DET1 effects in the light. DtRE binds two distinct activities in Arabidopsis seedling extracts: a novel activity with binding site CAAAACGC that we have named CAB2 DET1-associated factor 1 plus an activity that is likely to be the myb transcription factor Circadian Clock-Associated 1. Both activities are altered in dark-grown det1 extracts as compared with wild type, correlating a change in extractable DNA binding activity with a major change in CAB2 expression. We conclude that DET1 represses the CAB2 promoter in the dark by regulating the binding of two factors, CAB2 DET1-associated factor 1 and Circadian Clock-Associated 1, to the DtRE.
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