《植物生理学报》 2018, 54(1): 92-102

拟南芥AtMSI1蛋白与组蛋白去乙酰化酶AtHDA6的相互作用分析

袁连玉¹ , 梁国鲁², 杨松光³, 童华荣^1,*

¹西南大学食品科学学院, 重庆400715; ²西南大学园艺园林学院, 重庆400715; ³中国科学院华南植物园农业及资源植物研究中心, 广州510650

通信作者：童华荣；E-mail: huart@swu.edu.cn

摘　要：

表观遗传修饰是真核生物基因转录调控的重要方式, 在拟南芥生长发育过程中起着重要的作用。其中, PRC2 (Poly-comb Repressive Complex 2)蛋白复合体和组蛋白脱乙酰化酶HDAC均为重要的表观遗传调控蛋白。本研究利用酵母双杂交系统证明了拟南芥PRC2蛋白复合体成员中的AtMSI1 (Multi-Copy Suppressor of IRA1)蛋白和组蛋白脱乙酰化酶AtHDA6 (Histone Deacetylase 6)蛋白间的相互作用, 且确定其作用位点为AtMSI1蛋白的N端(1~114 aa)和C端(404~424 aa)的非特异性位点与AtHDA6蛋白的HD保守结构域 (27~332 aa)和N端非保守的结构域(1~26 aa)。本研究构建AtMSI1-YC和AtHDA6-YN融合蛋白表达载体, 共同转化拟南芥原生质体, 用双分子荧光互补(BiFC)技术验证了AtMSI1-YC和AtHDA6-YN蛋白间的相互作用, 并明确该相互作用的位点为细胞核。另外, pull-down试验再次证明了该相互作用的存在, 原核表达并纯化的AtMSI1-GST融合蛋白可以与AtHDA6-His重组蛋白发生体外结合。综上所述, 拟南芥AtMSI1与AtHDA6蛋白间存在相互作用, 且每个蛋白中均有2个特异性结合位点参与该相互作用。

关键词：拟南芥; AtMSI1; AtHDA6; 相互作用蛋白

收稿：2017-07-14   修定：2017-12-29

资助：国家自然科学基金(31400583)和重庆市基础与前沿研究计划一般项目(cstc2014jcyjA80011)。

Analysis of the interaction between AtMSI1 and AtHDA6 in Arabidopsis thaliana

YUAN Lian-Yu¹, LIANG Guo-Lu², YANG Song-Guang³, TONG Hua-Rong^1,*

¹College of Food Science, Southwest University, Chongqing 400715, China; ²College of Horticulture and Landscape Architecture, Southwest University, Chongqing 400715, China; ³Research Group of Plant Epigenetics, South China Botanical Garden, Chinese Academy of Sciences, Guangzhou 510650, China

Corresponding author: TONG Hua-Rong; E-mail: huart@swu.edu.cn

Abstract:

Chromatin modification is an effective way of transcriptional regulation in eukaryotes. Histone deacetylases and subuints of PRC2 (Poly-comb Repressive Complex 2) complexes are all essential for chromatin modification, and they play important roles in growth and development regulation in Arabidopsis thaliana. The yeast two-hybrid assay indicated that AtMSI1 could interact with AtHDA6, and the interaction was further confirmed using bimolecular fluorescence complementation (BiFC) method and pull-down technology, which happened in the nucleus. Furthermore, the specific binding sites of these two proteins interaction were confirmed, which were the N terminal (1–114 aa) and C (404–424 aa) tail of AtMSI1 protein and the HD conserved domain (27–332 aa) and N-terminal (1–26 aa) specific domain of AtHDA6 protein respectively. Taken together, our results indicated that the AtMSI1 protein could directly interact with the histone deacetylase AtHDA6, and there were two specific sites in each protein involved in the binding process of this interaction in A. thaliana.

Key words: Arabidopsis thaliana; AtMSI1; AtHDA6; interacting protein