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 GmMYB173的亚细胞定位情况,发现 GmMYB173在细胞核中特异表达；序列分析发现GmMYB173与GmMYB176相似,具有1个14-3-3蛋白的潜在结合结构域,即pST结合结构域。通过重叠延伸PCR (SOE-PCR)删除了GmMYB173序列中pST结合结构域编码序列,发现GmMYB173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GmSGF14a~GmSGF14p均能与 GmMYB173互作。β-半乳糖苷酶活性分析发现,与 GmMYB173互作最强的是 GmSGF14n,其次是 GmSGF14k、GmSGF14e 和GmSGF14o。这些结果说明14-3-3蛋白不仅与GmMYB173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GmMYB173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 pST binding site, in GmMYB173 similar to that in GmMYB176. The nuclear-specific expression of GmMYB173 disappeared when the sequence of the pST binding site was deleted through the splicing by overlap extension PCR. All 14-3-3 proteins from GmSGF14a to GmSGF14p could interact with GmMYB173. Among them, GmSGF14n interacted with GmMYB173 strongest, GmSGF14e and GmSGF14k took second place, which was proved by theβ-galactosidase activity analysis. These results suggest that 14-3-3 proteins not only interact with GmMYB173, but also probably regulate its subcellular location. The information pro-vided by this study will facilitate the study of interaction relationship between 14-3-3 proteins and GmMYB173 and its function on the soybean development.
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