摘要
Arabidopsis cryptochrome 2 （CRY2） is a blue light receptor that mediates light inhibition of hypocotyl elongation and long-day promotion of floral initiation, CRY2 is known to undergo blue light-dependent phosphorylation, which is believed to serve regulatory roles in the function of CRY2. We report here on a biochemical and genetics study of CRY2 phosphorylation. Using mass spectrometry analysis, we identified three serine residues in the CCE domain of CRY2 （S588, S599, and S605） that undergo blue light-dependent phosphorylation in Arabidopsis seedlings. A study of serine-substitution mutations in the CCE domain of CRY2 demonstrates that CRY2 contains two types of phosphorylation in the CCE domain, one in the serine cluster that causes electrophoretic mobility upshift and the other outside the serine cluster that does not seem to cause mobility upshift. We showed that mutations in the serine residues within and outside the serine cluster diminished blue light-dependent CRY2 phosphorylation, degradation, and physiological activities. These results support the hypothesis that blue light-dependent phosphorylation of the CCE domain determines the photosensitivity of Arabidopsis CRY2.
Arabidopsis cryptochrome 2 （CRY2） is a blue light receptor that mediates light inhibition of hypocotyl elongation and long-day promotion of floral initiation, CRY2 is known to undergo blue light-dependent phosphorylation, which is believed to serve regulatory roles in the function of CRY2. We report here on a biochemical and genetics study of CRY2 phosphorylation. Using mass spectrometry analysis, we identified three serine residues in the CCE domain of CRY2 （S588, S599, and S605） that undergo blue light-dependent phosphorylation in Arabidopsis seedlings. A study of serine-substitution mutations in the CCE domain of CRY2 demonstrates that CRY2 contains two types of phosphorylation in the CCE domain, one in the serine cluster that causes electrophoretic mobility upshift and the other outside the serine cluster that does not seem to cause mobility upshift. We showed that mutations in the serine residues within and outside the serine cluster diminished blue light-dependent CRY2 phosphorylation, degradation, and physiological activities. These results support the hypothesis that blue light-dependent phosphorylation of the CCE domain determines the photosensitivity of Arabidopsis CRY2.
基金
This work is supported in part by the National Institute of Health （GM56265 to C.L., GM089778 to J.A.W.）, research funds from Fujian Agriculture and Forestry University （to the Basic Forestry and Proteomics Research Center）, Jilin University （research support to the Laboratory of Soil and Plant Molecular Genetics）, the MOA Transgenic Research Grant （2010ZX08010-002 to B.L.）, and the National Natural Science Foundation of China （31171176 to X.Y. and 31422041 to B.L.）.ACKNOWLEDGMENTS No conflict of interest declared.
