Kaiserli Eirini, Jenkins Gareth I
Plant Science Group, Division of Biochemistry and Molecular Biology, Institute of Biomedical and Life Sciences, University of Glasgow, Glasgow G12 8QQ, United Kingdom.
Plant Cell. 2007 Aug;19(8):2662-73. doi: 10.1105/tpc.107.053330. Epub 2007 Aug 24.
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.
拟南芥紫外线抗性位点8(UVR8)是一种特异性响应UV-B的信号转导元件,它与染色质结合,并通过协调一系列基因的表达来调节紫外线防护。在此,我们研究了UV-B如何调控UVR8。我们发现UV-B能刺激绿色荧光蛋白(GFP)-UVR8融合蛋白和天然UVR8的核积累。核积累导致UV-B诱导HY5基因表达,HY5基因编码UVR8信号通路中的一个关键转录效应因子。UVR8的核积累对UV-B具有特异性,在低光通量率下即可发生,并且在UV-B照射后5分钟内就能观察到。在GFP-UVR8上连接一个核输出信号(NES)会使其在没有UV-B的情况下定位于细胞质。然而,UV-B能促进NES-GFP-UVR8快速核积累,这表明存在一种协同的核转运机制。缺失N端23个氨基酸的UVR8在核转运方面存在缺陷。在UVR8上连接一个核定位信号(NLS)会使其组成性定位于细胞核。然而,NLS-GFP-UVR8仅在UV-B照射后才能赋予HY5基因表达,这表明核定位虽然是UVR8发挥功能所必需的,但不足以引发靶基因的表达;还需要UV-B来刺激细胞核中UVR8的功能。这些发现为UV-B调节植物基因表达的机制提供了新的见解。