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. 2013 Nov 25;110(50):20326–20331. doi: 10.1073/pnas.1314336110

Fig. 2.

Fig. 2.

UVR8W285A appears as a constitutive monomer and constitutively interacts with COP1 in transgenic plants. (A) Four-day-old seedlings were irradiated for 15 min with (+) or without (−) supplementary broadband UV-B. The dimer was observed on Western blots from SDS-PAGE without sample boiling (Upper), and the total amount of UVR8 protein was detected in samples denatured by boiling (Lower). UVR8 protein levels of wild-type (Ws) plants are compared with those in Pro35S::UVR8W285A lines #4 and #24 (W285A #4 and #24) (Upper), and UVR8 protein levels of a line overexpressing Pro35S::UVR8 (UVR8-Ox) are compared with levels in lines overexpressing Pro35S::UVR8W285A (#6 and #12, Lower). uvr8-7 is shown as negative control demonstrating the specificity of the detected bands. (B) UVR8W285A does not form homodimers in vivo in yeast two-hybrid growth assays, in contrast to UVR8 and UVR8W285F. (C) UVR8W285A interacts with COP1 in a UV-B–independent manner, in contrast to wild-type UVR8, whereas UVR8W285F shows no interaction with COP1. (D) UVR8W285A interacts with the 340 C-terminal amino acids of COP1 in a UV-B–independent manner in a yeast two-hybrid growth assay. EV, empty vector control.