Reconstituting Arabidopsis CRY2 Signaling Pathway in Mammalian Cells Reveals Regulation of Transcription by Direct Binding of CRY2 to DNA

Abstract

Summary: In response to blue light, cryptochromes photoexcite and interact with signal partners to transduce signal almost synchronously in plants. The detailed mechanism of CRY-mediated light signaling remains unclear: the photobiochemical reactions of cryptochrome are transient and synchronous, thus making the monitoring and analysis of each step difficult in plant cells. In this study, we reconstituted the Arabidopsis CRY2 signaling pathway in mammalian cells and investigated the biological role of Arabidopsis CRY2 in this heterologous system, eliminating the interferences of other plant proteins. Our results demonstrated that, besides being the light receptor, Arabidopsis CRY2 binds to DNA directly and acts as a transcriptional activator in a blue-light-enhanced manner. Similar to classic transcription factors, we found that the transcriptional activity of CRY2 is regulated by its dimerization and phosphorylation. In addition, CRY2 cooperates with CIB1 to regulate transcription by enhancing the DNA affinity and transcriptional activity of CIB1 under blue light. : Cryptochromes are the blue light receptors in plants. Yang et al. reconstitute the Arabidopsis thaliana CRY2 signaling pathway to investigate the mechanism of CRY2 transcriptional regulation in mammalian cells. They demonstrate that, besides being the light receptor, CRY2 binds DNA directly to activate transcription of FT in a blue-light-enhanced manner. Keywords: cryptochrome, transcriptional activity, DNA binding, phosphorylation, blue light specificity