Multiple DNA-Protein Complexes at a Circadian-Regulated Promoter Element

Higher plant CAB genes encode chlorophyll a/b binding proteins that are part of light-harvesting complexes in chloroplasts. Transcription of the Arabidopsis CAB2 (lhcb1*1) gene is under the control of a circadian oscillator and exhibits high amplitude diurnal oscillations that persist within a period close to 24 hr in the absence of environmental time cues. Initial deletion studies in transgenic tobacco have demonstrated that the region between -111 and -38 of the CAB2 promoter sequence confers circadian regulation to a luciferase (luc) reporter gene. We dissected this element further and characterized five DNA binding complexes from Arabidopsis whole-cell extracts that bind within this region of the promoter and may be components of the signal transduction pathway for the control of transcription by the circadian clock. The in vivo analysis of cab2::luc fusion constructs in transgenic Arabidopsis demonstrated that a circadian-regulated element lies within a 36-bp sequence that overlaps a conserved CCAAT box and contains binding sites for three putative transcription factors.