Using HEK293T Expression System to Study Photoactive Plant Cryptochromes

Liang Yang; Xu Wang; Weixian Deng; Weiliang Mo; Jie Gao; Qing Liu; Chuanyu Zhang; Qin Wang; Chentao Lin; Zecheng Zuo

doi:10.3389/fpls.2016.00940

Using HEK293T Expression System to Study Photoactive Plant Cryptochromes

Front Plant Sci. 2016 Jun 27:7:940. doi: 10.3389/fpls.2016.00940. eCollection 2016.

Authors

Liang Yang¹, Xu Wang², Weixian Deng¹, Weiliang Mo³, Jie Gao³, Qing Liu⁴, Chuanyu Zhang⁴, Qin Wang⁵, Chentao Lin⁶, Zecheng Zuo¹

Affiliations

¹ Laboratory of Soil and Plant Molecular Genetics, College of Plant Science, Jilin UniversityChangchun, China; Basic Forestry and Proteomics Research Center, Haixia Institute of Science and Technology, Fujian Agriculture and Forestry UniversityFuzhou, China.
² Basic Forestry and Proteomics Research Center, Haixia Institute of Science and Technology, Fujian Agriculture and Forestry UniversityFuzhou, China; Department of Molecular, Cell and Developmental Biology, University of California, Los AngelesCA, USA.
³ Laboratory of Soil and Plant Molecular Genetics, College of Plant Science, Jilin University Changchun, China.
⁴ Basic Forestry and Proteomics Research Center, Haixia Institute of Science and Technology, Fujian Agriculture and Forestry University Fuzhou, China.
⁵ Laboratory of Soil and Plant Molecular Genetics, College of Plant Science, Jilin UniversityChangchun, China; Department of Molecular, Cell and Developmental Biology, University of California, Los AngelesCA, USA.
⁶ Department of Molecular, Cell and Developmental Biology, University of California, Los Angeles CA, USA.

Abstract

Cryptochromes are photolyase-like blue light receptors that are conserved in plants and animals. Although the light-dependent catalytic mechanism of photolyase is well studied, the photochemical mechanism of cryptochromes remains largely unknown. Lack of an appropriate protein expression system to obtain photochemically active cryptochrome holoproteins is a technical obstacle for the study of plant cryptochromes. We report here an easy-to-use method to express and study Arabidopsis cryptochrome in HEK293T cells. Our results indicate that Arabidopsis cryptochromes expressed in HEK293T are photochemically active. We envision a broad use of this method in the functional investigation of plant proteins, especially in the large-scale analyses of photochemical activities of cryptochromes such as blue light-dependent protein-protein interactions.

Keywords: HEK293T; cryptochrome; expression system; linear DNA; photo-biochemical activity.