Bellizzi Anna, White Martyn K, Wollebo Hassen S
a Department of Neuroscience; Center for Neurovirology; Temple University School of Medicine ; Philadelphia , PA USA.
Cell Cycle. 2015;14(13):2075-9. doi: 10.1080/15384101.2015.1042631. Epub 2015 May 27.
Endoplasmic reticulum (ER) stress is caused by the accumulation of misfolded or unfolded proteins in the lumen of the endoplasmic reticulum. CCAAT/enhancer binding proteins are one of the cellular proteins whose expression is upregulated during ER stress. Previously, we have identified C/EBPbeta isoforms, especially LIP, as a negative regulator of polyomavirus JC (JCV), the causative agent of the demyelinating disease progressive multifocal leukoencephalopathy (PML). Here, we show that the induction of ER stress by thapsigargin increase the expression of endogenous LIP and the degradation of JCV T-antigen in a JCV-transgenic mouse tumor cell line. Our results also revealed that overexpression of LIP significantly reduced the level of T-Ag and this effect is reversed upon siRNA-mediated silencing of LIP. Immunoprecipitation/Western blot experiments indicated that LIP interacts with T-antigen directly. Treatment of cells that overexpress LIP with MG115, a proteasome inhibitor, partially rescued LIP-mediated degradation of T-antigen. Our observations point to a role of LIP in ER stress regulation of T-antigen stability and may open a new avenue to study host-virus interaction during ER stress.
内质网(ER)应激是由内质网腔中错误折叠或未折叠蛋白质的积累引起的。CCAAT/增强子结合蛋白是在内质网应激期间表达上调的细胞蛋白之一。此前,我们已确定C/EBPβ异构体,尤其是LIP,作为多瘤病毒JC(JCV)的负调节因子,JCV是脱髓鞘疾病进行性多灶性白质脑病(PML)的病原体。在此,我们表明毒胡萝卜素诱导的内质网应激会增加内源性LIP的表达,并在JCV转基因小鼠肿瘤细胞系中导致JCV T抗原的降解。我们的结果还显示,LIP的过表达显著降低了T-Ag的水平,而在siRNA介导的LIP沉默后,这种效应被逆转。免疫沉淀/蛋白质印迹实验表明LIP直接与T抗原相互作用。用蛋白酶体抑制剂MG115处理过表达LIP的细胞,部分挽救了LIP介导的T抗原降解。我们的观察结果表明LIP在内质网应激调节T抗原稳定性中起作用,并可能为研究内质网应激期间的宿主-病毒相互作用开辟一条新途径。