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

机译:UV-B促进拟南芥UV-B特定信号传导成分UVR8的快速核转运并激活其在细胞核中的功能。

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摘要

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.
机译:拟南芥抗紫外线LOCUS8(UVR8)是一种UV-B特异的信号传导组分,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的保护。在这里,我们研究了UV-B如何调节UVR8。我们显示,UV B刺激绿色荧光蛋白(GFP)-UVR8融合和天然UVR8的核积累。核积累导致HY5基因的UV-B诱导,编码UVR8途径的关键转录效应子。 UVR8的核积累是UV-B特有的,以低通量率发生,并且在UV-B暴露5分钟内即可观察到。在没有UV-B的情况下,将核输出信号(NES)附着到GFP-UVR8会引起胞质定位。但是,UV-B促进了NES-GFP-UVR8的快速核积累,表明核转运的协调机制。缺少N端23个氨基酸的UVR8在核转运中受损。核定位信号(NLS)附着到UVR8会导致本构核定位。但是,NLS-GFP-UVR8仅在UV-B照射后才赋予HY5基因表达,表明核定位(尽管是UVR8功能所必需的)不足以引起靶基因的表达。另外还需要UV-B来刺激细胞核中的UVR8功能。这些发现为紫外线B调节植物基因表达的机制提供了新的见解。

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