染色质阅读器蛋白EBS和SHL的诱导表达和体外SUMO化修饰鉴定  

Induced Expression and in Vitro SUMOylation Identification of Chromatin Reader Proteins EBS and SHL

作　　者：魏建 李肇敏 苏树杰 吴子熙 李梦瑶[1] 韩永峰[1] Wei Jian;Li Zhaomin;Su Shujie;Wu Zixi;Li Mengyao;Han Yongfeng(Hebei Normal University,Shijiazhuang 050024,China)

机构地区：[1]河北师范大学,河北石家庄050024

出　　处：《廊坊师范学院学报（自然科学版）》2024年第4期41-48,共8页Journal of Langfang Normal University(Natural Science Edition)

基　　金：河北省自然科学基金项目(C2022205017);河北师范大学博士基金项目(L2019B23)。

摘　　要：EBS和SHL是植物特有的一类染色质阅读器蛋白,通过特异性识别并结合两类功能相互拮抗的组蛋白修饰调控靶基因表达和相关生物学过程,但目前未见有关翻译后修饰调控其蛋白功能的报道。利用大肠杆菌SUMO化修饰重建体系发现,EBS和SHL都存在SUMO化修饰,且不论以哪种SUMO分子亚型为供体都只有一条SUMO化条带,根据SUMO化条带和与本底蛋白分子量差值推测均为单位点的单SUMO化修饰。进一步利用定点突变实验证实,K216是EBS唯一且关键的SUMO化位点,而SHL的SUMO化位点有待继续研究,但至少可以排除K111、K178和K220三个位点,为下一步探讨SUMO化修饰调控EBS和SHL的蛋白功能奠定了基础。EBS and SHL are plant-specific chromatin reader proteins,which regulate target gene expression and related biological processes by specifically recognizing and binding to two antagonistic histone modifications.However,there have been no reports on the regulation of their protein functions by post-translational modifications.In this study,after the E.coli SUMOylation reconstruction system is used,it is found that both EBS and SHL have SUMOylation,and there is only one SUMOylation band,no matter which SUMO molecular subtype is used as the donor.According to the difference of molecular weight between the SUMOylation band and the background protein band,it is inferred that both EBS and SHL are single-site single SUMOylation.Further site-directed mutagenesis experiments confirm that K216 is the single critical SUMOylation site for EBS,while the SUMOylation site for SHL remains to be further explored,but at least K111,K178,and K220 can be excluded.This study lays a foundation for further exploration of the regulation of EBS and SHL protein functions by SUMOylation modification.

关 键 词：染色质阅读器蛋白 EBS SHL SUMO化修饰 SUMO 

分 类 号：Q943[生物学—植物学]