The POZ/BTB protein NAC1 interacts with two different histone deacetylases in neuronal-like cultures
The POZ/BTB protein NAC1 interacts with two different histone deacetylases in neuronal-like cultures
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DOI：
10.1111/j.1471-4159.2005.03206.x
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发表时间：
2005-08-01
影响因子：
4.7
通讯作者：
Mackler, SA
中科院分区：
文献类型：
--
作者：
Korutla, L;Wang, PJ;Mackler, SA
NAC1是一种受可卡因调控的POZ/BTB（痘病毒锌指结构域/矮脚鸡、电车轨道、广泛复合物）蛋白。在对药物成瘾至关重要的中枢神经系统区域——伏隔核中，NAC1会因可卡因而选择性增加。然而，NAC1在细胞中的作用尚不清楚。两种NAC1异构体，即sNAC1（短型NAC1）和lNAC1（长型NAC1），可能作为其他POZ/BTB蛋白的辅阻遏物。本研究探讨了sNAC1和lNAC1是否与其他辅阻遏物存在蛋白质 - 蛋白质相互作用。组蛋白去乙酰化酶（HDAC）抑制剂可逆转sNAC1和lNAC1对Gal4荧光素酶的抑制作用，但仅在类神经元培养物中如此。由于这些抑制剂无法区分组蛋白去乙酰化酶，因此选择了两种组蛋白去乙酰化酶进行进一步研究。HDAC3和HDAC4均与sNAC1和lNAC1存在蛋白质 - 蛋白质相互作用。这通过免疫共沉淀、谷胱甘肽 - S - 转移酶（GST）下拉实验以及哺乳动物双杂交实验得以证明。重要的是，POZ结构域或无POZ结构域的NAC1都能与这两种HDAC结合。其他辅阻遏物，特别是NCoR（核受体辅阻遏物）、SMRT（视黄酸和甲状腺激素受体的沉默中介体）和mSin3a，与sNAC1和lNAC1不存在蛋白质 - 蛋白质相互作用。在GST下拉实验或哺乳动物双杂交实验中均未显示出蛋白质 - 蛋白质相互作用。综上所述，这些实验结果表明sNAC1和lNAC1募集组蛋白去乙酰化酶进行转录抑制，进一步增强了POZ/BTB蛋白介导的抑制作用。
NAC1 is a cocaine-regulated POZ/BTB (Pox virus and Zinc finger/Bric-a-brac Tramtrack Broad complex) protein. NAC1 is increased by cocaine selectively in the nucleus accumbens, a CNS region important for drug addiction. NAC1's role in the cell, however, is not known. Each of the two NAC1 isoforms, sNAC1 (short NAC1) and INAC1 (long NAC1), may serve as corepressors for other POZ/BTB proteins. This study investigated whether sNAC1 and INAC1 demonstrated protein-protein interactions with other corepressors. Histone deacetylase (HDAC) inhibition reversed sNAC1 and INAC1 repression of Gal4 luciferase, but only in neuronal-like cultures. Because these inhibitors do not distinguish among histone deacetylases, two histone deacetylases were selected for further study. HDAC 3 and 4 both demonstrated protein-protein interactions with sNAC1 and INAC1. This was shown using coimmunoprecipitations, glutathione-S-transferase (GST) pullclowns and mammalian two-hybrids. Importantly, either the POZ domain or NAC1 without the POZ domain can bind these two HDACs. Other corepressors, specifically NCoR (nuclear receptor corepressor), SMRT (silencing mediator for retinoid and thyroid hormone receptor) and mSin3a, do not exhibit protein-protein interactions with sNAC1 and INAC1. None showed protein-protein interactions in GST pullclowns or mammalian two-hybrids. Taken together, the results of these experiments indicate sNAC1 and INAC1 recruit histone deacetylases for transcriptional repression, further enhancing POZ/BTB protein mediated repression.