Wu Keqiang, Tian Lining, Zhou Changhe, Brown Daniel, Miki Brian
Department of Biology, West Virginia University, Morgantown 26506, USA.
Plant J. 2003 Apr;34(2):241-7. doi: 10.1046/j.1365-313x.2003.01714.x.
The four HD2 proteins of Arabidopsis thaliana (AtHD2A-D) belong to a unique class of histone deacetylases that is plant specific. Previously, we have demonstrated that one of the members, AtHD2A, can mediate transcriptional repression when targeted to the promoter of a reporter gene. Here, we report that AtHD2B and AtHD2C can also repress gene expression. AtHD2A and AtHD2C differ from AtHD2B and AtHD2D in the composition of their structural domains. Our data show that both structural types play a role in the repression of gene transcription. We demonstrate that AtHD2A can mediate gene repression through interactions with transcription factors in plants. By fusing AtHD2A with the DNA-binding domain of the plant transcriptional factor Pti4, the expression of a GCC box containing reporter gene was repressed. We also demonstrated repression of a GUS gene with GAL4 enhancers using transgenic plants that expressed a GAL4/AtHD2A fusion gene. Furthermore, the expression of the GAL4/AtHD2A protein using the seed-specific napin promoter (NAP2) and the constitutive tCUP promoter demonstrated that repression of transgenes could be achieved in a tissue-specific or unrestricted manner. Targeting of HD2 proteins to specific promoters using transcription factor DNA-binding domains may therefore provide a new technology for silencing target genes and pathways in plants as well as for assessing the function of unknown transcription factors.
拟南芥的四种HD2蛋白(AtHD2A - D)属于一类独特的组蛋白去乙酰化酶,是植物特有的。此前,我们已经证明其中一个成员AtHD2A,当靶向报告基因的启动子时可以介导转录抑制。在此,我们报道AtHD2B和AtHD2C也能抑制基因表达。AtHD2A和AtHD2C在其结构域组成上与AtHD2B和AtHD2D不同。我们的数据表明,这两种结构类型在基因转录抑制中都起作用。我们证明AtHD2A可以通过与植物中的转录因子相互作用来介导基因抑制。通过将AtHD2A与植物转录因子Pti4的DNA结合结构域融合,含有GCC盒的报告基因的表达受到抑制。我们还使用表达GAL4/AtHD2A融合基因的转基因植物,利用GAL4增强子证明了对GUS基因的抑制。此外,使用种子特异性的napin启动子(NAP2)和组成型的tCUP启动子表达GAL4/AtHD2A蛋白表明,可以以组织特异性或无限制的方式实现对转基因的抑制。因此,利用转录因子DNA结合结构域将HD2蛋白靶向特定启动子,可能为沉默植物中的靶基因和途径以及评估未知转录因子的功能提供一种新技术。