UV-B Promotes Rapid Nuclear Translocation of the Arabidopsis UV-B–Specific Signaling Component UVR8 and Activates Its Function in the Nucleus

Abstract

Arabidopsis thaliana UV RESISTANCE LOCUS8 (UVR8) is a UV-B–specific signaling component that binds to chromatin and regulates UV protection by orchestrating expression of a range of genes. Here, we studied how UV-B regulates UVR8. We show that UV-B stimulates the nuclear accumulation of both a green fluorescent protein (GFP)-UVR8 fusion and native UVR8. Nuclear accumulation leads to UV-B induction of the HY5 gene, encoding a key transcriptional effector of the UVR8 pathway. Nuclear accumulation of UVR8 is specific to UV-B, occurs at low fluence rates, and is observed within 5 min of UV-B exposure. Attachment of a nuclear export signal (NES) to GFP-UVR8 causes cytosolic localization in the absence of UV-B. However, UV-B promotes rapid nuclear accumulation of NES-GFP-UVR8, indicating a concerted mechanism for nuclear translocation. UVR8 lacking the N-terminal 23 amino acids is impaired in nuclear translocation. Attachment of a nuclear localization signal (NLS) to UVR8 causes constitutive nuclear localization. However, NLS-GFP-UVR8 only confers HY5 gene expression following UV-B illumination, indicating that nuclear localization, although necessary for UVR8 function, is insufficient to cause expression of target genes; UV-B is additionally required to stimulate UVR8 function in the nucleus. These findings provide new insights into the mechanisms through which UV-B regulates gene expression in plants.