小麦翻译控制肿瘤蛋白TCTP与蔗糖非酵解型蛋白激酶SnRK1的相互作用

Interaction between wheat translationally controlled tumor protein TCTP and SNF1-related protein kinase SnRK1

摘要:

翻译控制肿瘤蛋白 (Translationally controlled tumor protein, TCTP) 广泛存在于真核细胞中，参与调节细胞分裂、植物生长发育，并介导植物抵御病原物侵染。蔗糖非酵解型蛋白激酶 (SNF1- related protein kinase, SnRK1)在酵母、动物和植物中非常保守，并参与包括糖代谢和抵抗非生物和生物胁迫在内的一系列生理过程。本实验室前期工作证明TaTCTP响应叶锈菌侵染并参与诱发寄主产生防卫反应。为了深入探讨TaTCTP在叶锈菌侵染小麦诱发的防卫反应中发挥的作用，采用串联亲和纯化 (TAP) 与质谱 (MS) 联用技术，鉴定出SnRK1可能为TaTCTP潜在互作蛋白。文中对TCTP和SnRK1的相互作用进行了研究。酵母双杂交结果表明，同时携带TCTP和SnRK1的酵母可以在SD/-Leu/-Trp/-His/-Ade (SD/-LWHA, 四缺) 培养基上生长，说明TCTP与SnRK1在酵母双杂交系统中可以发生相互作用；通过双分子荧光互补实验，发现TCTP与SnRK1发生相互作用的荧光信号分布在细胞质中；进一步用Co-IP实验证明TCTP和SnRK1可以发生相互作用。本研究为深入研究TaTCTP在小麦与叶锈菌互作过程中的作用机制奠定了重要基础，对进一步完善小麦抵御叶锈菌侵染的分子机理具有重要意义。

Abstract:

Translationally controlled tumor proteins (TCTP) and SNF1- related protein kinase (SnRK1) are conserved and widely present in eukaryotic cells. TCTP regulates cell division, plant growth and development, and mediates plant resistance against pathogen infection. SnRK1 participates in a range of physiological processes including sugar metabolism and resistance to abiotic and biotic stresses. Previous work in our laboratory demonstrated that wheat TCTP can respond to Puccinia triticina infection and induce host defense responses. In order to further investigate the mechanism of TaTCTP in wheat resistance to Puccinia triticina infection, we used TAP (tandem affinity purification) and mass spectrometry to screen the potential interactants of TaTCTP. A SNF1- related protein kinase (SnRK1) was identified as a potential interacting protein of TaTCTP. The results of yeast two-hybrid assay showed that TCTP could interact with SnRK1 in yeast, and the yeast carrying TCTP and SnRK1 could grow on SD/-Leu/-Trp/-His/-Ade (SD/-LWHA) medium. The fluorescence signal of the interaction between TCTP and SnRK1 was found to be distributed in the cytoplasm in the Bi-fluorescense complementation experiment. Co-IP experiments further showed that TCTP and SnRK1 could interact in plant cells. This study lays an important foundation for further studying the mechanism of TaTCTP in the interaction between wheat and Puccinia triticina, and it play a great influence on further improving the molecular mechanism of wheat resistant to Puccinia triticina.