Zhu Yan, Rong Liang, Luo Qiang, Wang Baihui, Zhou Nana, Yang Yue, Zhang Chi, Feng Haiyang, Zheng Lina, Shen Wen-Hui, Ma Jinbiao, Dong Aiwu
State Key Laboratory of Genetic Engineering, Collaborative Innovation Center of Genetics and Development, International Associated Laboratory of CNRS-Fudan-HUNAU on Plant Epigenome Research, Department of Biochemistry, Institute of Plant Biology, School of Life Sciences, Fudan University, Shanghai 20043, P.R. China
State Key Laboratory of Genetic Engineering, Collaborative Innovation Center of Genetics and Development, Department of Biochemistry, Institute of Plant Biology, School of Life Sciences, Fudan University, Shanghai 20043, P.R. China.
Plant Cell. 2017 Feb;29(2):260-276. doi: 10.1105/tpc.16.00719. Epub 2017 Jan 30.
NUCLEOSOME ASSEMBLY PROTEIN1 (NAP1) defines an evolutionarily conserved family of histone chaperones and loss of function of the NAP1 family genes () and causes abnormal root hair formation. Yet, the underlying molecular mechanisms remain unclear. Here, we show that NRP1 interacts with the transcription factor WEREWOLF (WER) in vitro and in vivo and enriches at the () promoter in a WER-dependent manner. Crystallographic analysis indicates that NRP1 forms a dimer via its N-terminal α-helix. Mutants of NRP1 that either disrupt the α-helix dimerization or remove the C-terminal acidic tail, impair its binding to histones and WER and concomitantly lead to failure to activate transcription and to rescue the mutant phenotype. Our results further demonstrate that WER-dependent enrichment of NRP1 at the promoter is involved in local histone eviction and nucleosome loss in vivo. Biochemical competition assays imply that the association between NRP1 and histones may counteract the inhibitory effect of histones on the WER-DNA interaction. Collectively, our study provides important insight into the molecular mechanisms by which histone chaperones are recruited to target chromatin via interaction with a gene-specific transcription factor to moderate chromatin structure for proper root hair development.
核小体组装蛋白1(NAP1)定义了一个进化上保守的组蛋白伴侣家族,NAP1家族基因()功能丧失会导致根毛形成异常。然而,其潜在的分子机制仍不清楚。在这里,我们表明NRP1在体外和体内与转录因子狼蛛(WER)相互作用,并以WER依赖的方式在()启动子处富集。晶体学分析表明,NRP1通过其N端α螺旋形成二聚体。破坏α螺旋二聚化或去除C端酸性尾巴的NRP1突变体,损害其与组蛋白和WER的结合,并随之导致无法激活转录以及无法挽救突变体表型。我们的结果进一步证明,NRP1在启动子处的WER依赖富集参与了体内局部组蛋白驱逐和核小体丢失。生化竞争试验表明,NRP1与组蛋白之间的结合可能抵消组蛋白对WER-DNA相互作用的抑制作用。总的来说,我们的研究为组蛋白伴侣通过与基因特异性转录因子相互作用被招募到目标染色质以调节染色质结构促进根毛正常发育的分子机制提供了重要见解。