Interaction of the Arabidopsis UV-B-specific signaling component UVR8 with chromatin
Interaction of the Arabidopsis UV-B-specific signaling component UVR8 with chromatin
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DOI：
10.1093/mp/ssm012
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发表时间：
2008-01-01
期刊：
影响因子：
27.5
通讯作者：
Jenkins, Gareth I.
中科院分区：
文献类型：
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作者：
Cloix, Catherine;Jenkins, Gareth I.
拟南芥抗紫外线基因座8 (UVR8) 是一种UV-B 特异性信号成分，可调节一系列与紫外线防护相关的基因的表达。在这里，我们研究了 UVR8 与染色质的相互作用。在野生型植物的染色质免疫沉淀 (ChIP) 测定中使用 UVR8 特异性抗体，我们发现天然 UVR8 在体内与染色质结合。使用抗 GFP 抗体对表达 GFP - UVR8 融合体的植物进行的类似实验表明，UVR8 与含有 HY5 基因的染色质相对较小的区域相关。 UVR8 与含有其他基因启动子区域的染色质相互作用，但不与其调节的所有基因相互作用。 UVR8 与染色质的相互作用不需要 UV-B，因为在没有 UV-B 的情况下观察到与多个基因位点的关联。 Pull-down 测定表明 UVR8 在体内与组蛋白结合，而竞争实验表明该相互作用优先与组蛋白 H2B 结合。使用识别特定组蛋白修饰的抗体进行 ChIP 实验表明，UV-B 刺激的某些基因转录可能与组蛋白修饰相关。特别是，ELIP1 启动子在 UV-B 暴露后显示出二乙酰组蛋白 H3 (K9/K14) 的显着富集。这些发现加深了对关键 UV-B 特异性调节因子 UVR8 与染色质相互作用的理解。
Arabidopsis UV RESISTANCE LOCUS8 (UVR8) is a UV-B-specific signaling component that regulates expression of a range of genes concerned with UV protection. Here, we investigate the interaction of UVR8 with chromatin. Using antibodies specific to UVR8 in chromatin immunoprecipitation (ChIP) assays with wild-type plants, we show that native UVR8 binds to chromatin in vivo. Similar experiments using an anti-GFP antibody with plants expressing a GFP - UVR8 fusion show that UVR8 associates with a relatively small region of chromatin containing the HY5 gene. UVR8 interacts with chromatin containing the promoter regions of other genes, but not with all the genes it regulates. UV-B is not required for the interaction of UVR8 with chromatin because association with several gene loci is observed in the absence of UV-B. Pull-down assays demonstrate that UVR8 associates with histones in vivo and competition experiments indicate that the interaction is preferentially with histone H2B. ChIP experiments using antibodies that recognize specific histone modifications indicate that the UV-B-stimulated transcription of some genes may be correlated with histone modification. In particular, the ELIP1 promoter showed a significant enrichment of diacetyl histone H3 (K9/K14) following UV-B exposure. These findings increase understanding of the interaction of the key UV-B-specific regulator UVR8 with chromatin.