泛素连接酶MDM2促进外源NOLC1的泛素化和降解

基础医学与临床 ›› 2014, Vol. 34 ›› Issue (3): 314-317.

泛素连接酶MDM2促进外源NOLC1的泛素化和降解

王小蓉¹,郭正光¹,高友鹤²

1. 中国医学科学院基础医学研究所
2. 中国医学科学院基础医学研究所北京协和医学院基础医学院

收稿日期:2013-09-26 修回日期:2013-12-14 出版日期:2014-03-05 发布日期:2014-02-27
通讯作者: 高友鹤 E-mail:gaoyouhe@gmail.com
基金资助:
国家基础研究计划(973);国家高技术研究发展计划（863）;高校长江学者创新研究团队;111计划

Ubiquitin ligase MDM2 promotes the ubiquitination and degradation of exogenous NOLC1

Received:2013-09-26 Revised:2013-12-14 Online:2014-03-05 Published:2014-02-27
Contact: GAO You-he E-mail:gaoyouhe@gmail.com

摘要/Abstract

摘要： 目的研究泛素连接酶MDM2对其调节因子NOLC1的泛素化和降解。方法克隆原核表达了NOLC1全长蛋白和其核定位信号区域，在体外泛素化体系中利用有泛素连接酶活性的重组MDM2研究其对NOLC1的泛素化。在哺乳动物细胞中，研究MDM2对外源转染的NOLC1的降解。结果在体外泛素化体系中MDM2泛素化NOLC1全长和其核定位信号区域；在哺乳动物细胞中，MDM2促进外源NOLC1降解超过70%，且NOLC1的降解通过蛋白酶体途经实现。结论 MDM2促进外源NOLC1的泛素化和降解，为研究MDM2-TP53-NOLC1之间的相互调节提供了新的线索。

关键词: 泛素连接酶, 泛素化, MDM2, NOLC1

Abstract: Objective To study the role of MDM2 in the ubiquitination of NOLC1, a regulator of MDM2. Methods Full-length NOLC1 proteins and its nuclear localization signal region were cloned and expressed in E.coli. Recombinant human MDM2 with ubiquitin ligase activity was used to study the ubiquitination of NOLC1 by MDM2 in in vitro ubiquitination system. The degradation of exogenous NOLC1 by MDM2 was studied in mammalian cells. Results MDM2 promoted the ubiquitination of full-length NOLC1 proteins and its Nuclear localization signal region in in vitro ubiquitination system. MDM2 promoted the proteasome dependent degradation of exogenous NOLC1 by over 70% in mammalian cells. Conclusion Ubiquitin ligase MDM2 promotes the ubiquitination and degradation of exogenous regulator NOLC1, which provides important clues for study the regulation of MDM2-TP53-NOLC1.

Key words: ubiquitin ligase, ubiquitination, MDM2, NOLC1

中图分类号:

[R34]

王小蓉郭正光高友鹤. 泛素连接酶MDM2促进外源NOLC1的泛素化和降解[J]. 基础医学与临床, 2014, 34(3): 314-317.