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Comparative properties and functions of type 2 and type 4 pigeon cryptochromes

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Abstract

Two types of vertebrate cryptochromes (Crys) are currently recognized. Type 2 Crys function in the molecular circadian clock as light-independent transcriptional repressors. Type 4 Crys are a newly discovered group with unknown function, although they are flavoproteins, and therefore, may function as photoreceptors. It has been postulated that Crys function in light-dependent magnetoreception, which is thought to contribute towards homing and migratory behaviors. Here we have cloned and annotated the full-length pigeon ClCry1, ClCry2, and ClCry4 genes, and characterized the full-length proteins and several site-directed mutants to investigate the roles of these proteins. ClCry1 and ClCry2 are phylogenetically grouped as Type 2 Crys and thus are expected to be core components of the pigeon circadian clock. Interestingly, we find that ClCry4 is properly annotated as a Type 4 Cry. It appears that many birds possess a Type 4 Cry which, as in pigeon, is misannotated. Like the Type 2 Crys, ClCry4 is widespread in pigeon tissues. However, unlike the Type 2 Crys, ClCry4 is cytosolic, and purified ClCry4 possesses FAD cofactor, which confers characteristic UV–Vis spectra as well as two photochemical activities. We find that ClCry4 undergoes light-dependent conformational change, which is a property of insect Type 1 Crys involved in the insect-specific pathway of photoentrainment of the biological clock. ClCry4 can also be photochemically reduced by a mechanism common to all FAD-containing Cry family members, and this mechanism is postulated to be influenced by the geomagnetic field. Thus pigeon Crys control circadian behavior and may also have photosensory function.

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Funding

This work is supported by the National Natural Science Foundation of China (Grant no. 21403298) and the China Specialized Research Fund for the Doctoral Program of Higher Education (Grant no. 20134307120015) to Jing Wang. This work was also supported by NIH Grants GM118102 and ES027255 to Aziz Sancar.

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Authors and Affiliations

  1. College of Science, National University of Defense Technology, Changsha, 410073, Hunan, People’s Republic of China

    Xuefeng Wang, Chengyu Jing, Wenjian Wu & Jing Wang

  2. Department of Biochemistry and Biophysics, University of North Carolina School of Medicine, Chapel Hill, NC, 27599, USA

    Xuefeng Wang, Christopher P. Selby, Yi-Ying Chiou, Yanyan Yang & Aziz Sancar

  3. Institute of Biochemistry, National Chung Hsing University, Taichung, Taiwan

    Yi-Ying Chiou

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  1. Xuefeng Wang
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  2. Chengyu Jing
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Correspondence to Aziz Sancar or Jing Wang.

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Wang, X., Jing, C., Selby, C.P. et al. Comparative properties and functions of type 2 and type 4 pigeon cryptochromes. Cell. Mol. Life Sci. 75, 4629–4641 (2018). https://doi.org/10.1007/s00018-018-2920-y

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