PHF6 Interacts with the Nucleosome Remodeling and Deacetylation (NuRD) Complex
PHF6 Interacts with the Nucleosome Remodeling and Deacetylation (NuRD) Complex
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DOI：
10.1021/pr3004369
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发表时间：
2012-08-01
影响因子：
4.4
通讯作者：
Picketts, David J.
中科院分区：
文献类型：
--
作者：
Todd, Matthew A. M.;Picketts, David J.
PHF 6中的突变是Borjeson-Forssman-Lehman综合征（BFLS），一种X连锁智力残疾（XLID）疾病，以及T细胞急性淋巴细胞白血病（T-ALL）和急性髓性白血病（AML）的原因。PHF 6基因编码具有两个植物同源结构域（PHD）样锌指结构域的蛋白。由于许多PHD样结构域的功能是靶向染色质重塑后修饰的组蛋白，这表明PHF 6在染色质调控中的作用。然而，PHD结构域通常与催化结构域相关，这是PHF 6缺乏的特征。这种独特的结构域结构及其细胞功能的最小信息促使我们进行蛋白质组学筛选，以确定PHF 6结合伴侣。我们在HEK 293 T细胞中表达了重组Flag标记的PHF 6，用于免疫共沉淀，并通过质谱分析纯化的产物。我们鉴定了参与核糖体生物发生、RNA剪接和染色质调节的蛋白质，这与PHF 6定位于核质和核仁一致。值得注意的是，PHF 6与核小体重塑和脱乙酰化（NuRD）复合物的多种成分共纯化，包括CHD 4、HDAC 1和RBBP 4。我们表明，PHF 6 NURD复合物不存在于核仁中，但仅限于核质。与NuRD的关联代表了PHF 6的第一个已知相互作用，并暗示它参与染色质调控。
Mutations in PHF6 are the cause of Borjeson-Forssman-Lehman syndrome (BFLS), an X-linked intellectual disability (XLID) disorder, and both T-cell acute lymphoblastic leukemia (T-ALL) and acute myeloid leukemia (AML). The PHF6 gene encodes a protein with two plant homeodomain (PHD)-like zinc finger domains. As many PHD-like domains function to target chromatin remodelers to post-translationally modified histones, this suggests a role for PHF6 in chromatin regulation. However, PHD domains are usually found in association with a catalytic domain, a feature that is lacking in PHF6. This distinct domain structure and the minimal information on its cellular function prompted us to perform a proteomic screen to identify PHF6 binding partners. We expressed recombinant Flag-tagged PHF6 in HEK 293T cells for coimmunoprecipitation, and analyzed the purified products by mass spectrometry. We identified proteins involved in ribosome biogenesis, RNA splicing, and chromatin regulation, consistent with PHF6 localization to both the nucleoplasm and nucleolus. Notably, PHF6 copurified with multiple constituents of the nucleosome remodeling and deacetylation (NuRD) complex, including CHD4, HDAC1, and RBBP4. We demonstrate that this PHF6 NuRD complex is not present in the nucleolus but is restricted to the nucleoplasm. The association with NuRD represents the first known interaction for PHF6 and implicates it in chromatin regulation.