利用酵母双杂交系统筛选水稻组蛋白去乙酰化酶HDA705的互作蛋白

Screening of Proteins Interacting with Histone Deacetylase HDA705 in Rice (Oryza sativa) by Yeast Two Hybrid System

摘要:

为了解水稻(Oryza sativa)组蛋白去乙酰化酶HDA705 的生物学功能,构建了HDA705 酵母双杂交诱饵表达载体与双杂交文库,并筛选了与HDA705 相互作用的蛋白。结果表明,HDA705 的诱饵载体无自激活活性且对酵母无毒性作用,文库的滴度也适合常规的酵母双杂交文库筛选。通过对酵母双杂交文库的筛选,共获得了164 个阳性克隆,经DNA 测序分析,这些克隆编码47 个可能与HDA705 相互作用的蛋白,其中包括3 个在逆境响应或激素信号转导过程中起到重要作用的(辅)转录因子、6 个参与光合作用的叶绿体蛋白、1 个含有R3H 结构域的蛋白以及22 种酶类等。这为进一步研究HDA705 的生物学功能提供了重要的线索。

Abstract:

In order to understand the biological function of histone deacetylase HDA705 in rice, the yeast twohybrid bait vector of HDA705 and the cDNA library were constructed and the interaction partners of HDA705 were screened. The results showed that HDA705 bait vector showed no self-activating and no toxicity to yeast cells. Furthermore, the titer of cDNA library was suitable for normal yeast two-hybrid screening assay. There were 164 positive clones by yeast two-hybrid screening. DNA sequencing analysis indicated that these positive clones encoded 47 proteins which may interact with HDA705. These proteins included 3 transcription factors (or transcription co-factors) which play key roles in stress responses and hormone signaling pathways, 6 chloroplast located proteins which were involved in photosynthesis, 1 protein containing the R3H domain structure, and 22 enzymes in plant, etc. These may provide important cues to further study the biological function of HDA705 in rice.