大豆14-3-3蛋白与转录因子蛋白GmMYB173的互作

Interaction of Soybean 14-3-3 Proteins with Transcription Factor Gm MYB173

14-3-3蛋白家族在真核生物中普遍存在,可与其他蛋白相互作用,调控多种生理生化过程。MYB基因家族作为植物中最大的一类转录因子,广泛参与了植物的生长发育和代谢调控。本研究通过分析1个从自贡冬豆中克隆的MYB转录因子Gm MYB173的亚细胞定位情况,发现Gm MYB173在细胞核中特异表达;序列分析发现Gm MYB173与Gm MYB176相似,具有1个14-3-3蛋白的潜在结合结构域,即p ST结合结构域。通过重叠延伸PCR(SOE-PCR)删除了Gm MYB173序列中p ST结合结构域编码序列,发现Gm MYB173细胞核表达特异性消失。酵母双杂交互作分析表明,大豆基因组中所有具有表达的16个14-3-3蛋白Gm SGF14a^Gm SGF14p均能与Gm MYB173互作。β-半乳糖苷酶活性分析发现,与Gm MYB173互作最强的是Gm SGF14n,其次是Gm SGF14k、Gm SGF14e和Gm SGF14o。这些结果说明14-3-3蛋白不仅与Gm MYB173互作,且可能调控其在细胞内的定位,有助于研究14-3-3蛋白与Gm MYB173的互作关系及其在大豆生长发育中的作用。

14-3-3 proteins, nearly existing in all eukaryotic cells, may regulate many physiological and biochemical processes through interacting with other proteins. As the largest class of transcription factors in plants, MYB gene family is widely involved in plant growth and metabolism regulation. A gene cloned from soybean cultivar Zigongdongdou, and specifically expressed in the nuclear in the subcellular location assay. Sequence analysis showed that there was a binding site of 14-4-4 proteins, namely the p ST binding site, in Gm MYB173 similar to that in Gm MYB176. The nuclear-specific expression of Gm MYB173 disappeared when the sequence of the p ST binding site was deleted through the splicing by overlap extension PCR. All 14-3-3 proteins from Gm SGF14 a to Gm SGF14 p could interact with Gm MYB173. Among them, Gm SGF14 n interacted with Gm MYB173 strongest, Gm SGF14 e and Gm SGF14 k took second place, which was proved by the β-galactosidase activity analysis. These results suggest that 14-3-3 proteins not only interact with Gm MYB173, but also probably regulate its subcellular location. The information provided by this study will facilitate the study of interaction relationship between 14-3-3 proteins and Gm MYB173 and its function on the soybean development.