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

Liang Yang; Weiliang Mo; Xiaolan Yu; Nan Yao; Zeng Zhou; Xiaolu Fan; Li Zhang; Mingxin Piao; Shiming Li; Dehong Yang; Chentao Lin; Zecheng Zuo

doi:10.1016/j.celrep.2018.06.069

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

Cell Rep. 2018 Jul 17;24(3):585-593.e4. doi: 10.1016/j.celrep.2018.06.069.

Authors

Liang Yang¹, Weiliang Mo², Xiaolan Yu³, Nan Yao¹, Zeng Zhou³, Xiaolu Fan³, Li Zhang², Mingxin Piao¹, Shiming Li³, Dehong Yang², Chentao Lin⁴, Zecheng Zuo⁵

Affiliations

¹ Basic Forestry and Proteomics Research Center, Fujian Agriculture and Forestry University, Fuzhou 350002, China; Jilin Province Engineering Laboratory of Plant Genetic Improvement, College of Plant Science, Jilin University, 5333 Xi'an Road, Changchun 130062, China.
² Jilin Province Engineering Laboratory of Plant Genetic Improvement, College of Plant Science, Jilin University, 5333 Xi'an Road, Changchun 130062, China.
³ Basic Forestry and Proteomics Research Center, Fujian Agriculture and Forestry University, Fuzhou 350002, China.
⁴ Department of Molecular, Cell, and Developmental Biology, University of California, Los Angeles, Los Angeles, CA 90095, USA.
⁵ Basic Forestry and Proteomics Research Center, Fujian Agriculture and Forestry University, Fuzhou 350002, China; Jilin Province Engineering Laboratory of Plant Genetic Improvement, College of Plant Science, Jilin University, 5333 Xi'an Road, Changchun 130062, China. Electronic address: zuozecheng@fafu.edu.cn.

PMID: 30021157
DOI: 10.1016/j.celrep.2018.06.069

Abstract

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.

Keywords: DNA binding; blue light specificity; cryptochrome; phosphorylation; transcriptional activity.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Arabidopsis / metabolism*
Arabidopsis / radiation effects
Arabidopsis Proteins / metabolism*
Cryptochromes / metabolism*
DNA / metabolism*
HEK293 Cells
Humans
Light
Light Signal Transduction / radiation effects
Phosphorylation / radiation effects
Protein Binding / radiation effects
Protein Multimerization / radiation effects
Signal Transduction*
Transcription, Genetic* / radiation effects
Transcriptional Activation / genetics

Substances

Arabidopsis Proteins
CRY2 protein, Arabidopsis
Cryptochromes
DNA

Grants and funding

R01 GM056265/GM/NIGMS NIH HHS/United States