Post-transcriptional regulation of dual-specificity phosphatase-1 by RNA-binding protein HuR T118 in heat shock
10.3969/j.issn.1673-4254.2014.06.03
- VernacularTitle:HuR蛋白118位苏氨酸对热应激状态下的双特异性磷酸酶1的转录后调控
- Author:
Chuanli ZHANG
1
;
Haihua LUO
;
Yong JIANG
Author Information
1. 南方医科大学病理生理教研室/广东省蛋白质组学重点实验室
- Keywords:
HuR protein;
dual-specificity phosphatase-1;
post-transcriptional regulation
- From:
Journal of Southern Medical University
2014;(6):766-770
- CountryChina
- Language:Chinese
-
Abstract:
Objective To investigate the post-transcriptional regulation of dual-specificity phosphatase-1 (DUSP1) by the RNA-binding protein HuR in heat shock. Methods The recombinant plasmids carrying wild-type (WT) HuR or its mutants at threonine 118 were constructed and transiently transfected into NIH 3T3 cells via liposome, and the changes in the expressions of DUSP1 mRNA and protein were detected by quantitative real-time PCR and Western blotting, respectively. Results Heat shock caused significantly enhanced phosphorylation of HuR at the residue T118. In 3T3 cells transfected with the plasmids carrying wild-type HuR for its over-expression showed significantly up-regulated DUSP1 mRNA and protein expressions at 24 h after transfection. Over-expression of HuR(T118A) down-regulated DUSP1 mRNA and protein expressions in cells challenged with heat shock, while HuR(T118E) over-expression significantly increased DISP1 expression at both mRNA and protein levels. After heat shock, HuR(WT) translocated from the cell nucleus to the cytoplasm to form particles. HuR(T118E) was diffusely distributed in the cytoplasm before heat shock and formed particles after heat shock. HuR(T118A) did not undergo such translocation in response to heat shock challenge. Conclusion HuR regulates DUSP1 mRNA and protein expression at the post-transcriptional level to increase its expression after heat shock by enhancing the phosphorylation HuR T118.
