The 5′UTR of CCA1 includes an autoregulatory cis element that segregates between light and circadian regulation of CCA1 and LHY

Abstract

The transcription factor CCA1 (CIRCADIAN CLOCK ASSOCIATED 1) participates in both light and circadian clock regulation in Arabidopsis. Two sets of transgenic plants in which GFP was fused to the CCA1 promoter with (1.3-kb fragment) or without (1.01-kb fragment) its 5'UTR were engineered. The transgenic plants transformed with the promoter including the 5'UTR had altered circadian regulation resulting in elongated hypocotyls, a bushy appearance and delayed flowering. In contrast, the transgenic plants transformed with the promoter without the 5'UTR showed earlier flowering than the wild type. Changes in CCA1, LHY and TOC1 gene expression were investigated under light-dark (L:D) fluctuations, continuous darkness (D:D) and continuous light (L:L). The circadian expression of CCA1 was altered in both sets of transgenic plants, being repressed in the plants transformed with the 1.01-kb fragment and constitutively overexpressed in those transformed with the 1.3-kb fragment. Under L:D conditions, regulation of LHY and TOC1 expression was separated from CCA1 regulation in both sets of transgenic plants, with intact rhythmic expression of both LHY and TOC1. Under D:D conditions, the rhythmic expression of LHY and TOC1 was lost in the 1.3 plants but retained with some erratic pattern under L:L conditions. In the 1.01 plants, under both D:D and L:L conditions the rhythmic expression was retained. These results indicate separate light-induced signal-transmission pathways for LHY and CCA1.