Paakinaho Ville, Kaikkonen Sanna, Levonen Anna-Liisa, Palvimo Jorma J
Institute of Biomedicine/Medical Biochemistry, University of Eastern Finland, Kuopio, Finland.
A. I. Virtanen Institute, University of Eastern Finland, Kuopio, Finland.
Mol Cell Biol. 2014 Sep;34(17):3202-13. doi: 10.1128/MCB.00748-14. Epub 2014 Jun 30.
Cortisol, the central stress hormone in humans, activates the glucocorticoid receptor (GR). Anti-inflammatory effects are the most important pharmaceutical effects mediated by the GR. Inasmuch as electrophilic cyclopentenone prostaglandin 15-deoxy-Δ(12,14)-prostaglandin J2 (15d-PGJ2) has potent anti-inflammatory properties and activates the SUMOylation pathway, we have investigated the effect of 15d-PGJ2 on glucocorticoid signaling and receptor SUMOylation. To this end, we studied isogenic HEK293 cells expressing either wild-type GR or SUMOylation-defective GR. Interestingly, 15d-PGJ2 triggered SUMO-2 and -3 (SUMO-2/3) modification in the primary SUMOylation sites of the GR. Gene expression profiling and pathway analyses indicate that 15d-PGJ2 inhibits GR signaling in a genome-wide fashion that is significantly dependent on the GR SUMOylation sites. Chromatin immunoprecipitation assays showed that the repressive effect of 15d-PGJ2 on GR target gene expression occurs in parallel with the inhibition of receptor binding to the target gene chromatin. Furthermore, depletion of UBC9, the sole SUMO E2 conjugase, from HEK293 cells confirmed the involvement of active SUMOylation in the regulatory process. Taken together, our data indicate that GR SUMOylation modulates the glucocorticoid signaling during acute cell stress. Our data also suggest that GR SUMOylation modulates cross talk of the glucocorticoid signaling with other transcription factors that are responsive to cell stress.
皮质醇是人类的主要应激激素,可激活糖皮质激素受体(GR)。抗炎作用是GR介导的最重要的药物效应。鉴于亲电环戊烯酮前列腺素15-脱氧-Δ(12,14)-前列腺素J2(15d-PGJ2)具有强大的抗炎特性并激活SUMO化途径,我们研究了15d-PGJ2对糖皮质激素信号传导和受体SUMO化的影响。为此,我们研究了表达野生型GR或SUMO化缺陷型GR的同基因HEK293细胞。有趣的是,15d-PGJ2在GR的主要SUMO化位点触发了SUMO-2和-3(SUMO-2/3)修饰。基因表达谱分析和通路分析表明,15d-PGJ2以全基因组方式抑制GR信号传导,这显著依赖于GR SUMO化位点。染色质免疫沉淀分析表明,15d-PGJ2对GR靶基因表达的抑制作用与受体与靶基因染色质结合的抑制同时发生。此外,从HEK293细胞中耗尽唯一的SUMO E2连接酶UBC9证实了活性SUMO化参与了调节过程。综上所述,我们的数据表明GR SUMO化在急性细胞应激期间调节糖皮质激素信号传导。我们的数据还表明,GR SUMO化调节糖皮质激素信号传导与其他对细胞应激有反应的转录因子之间的相互作用。