Joshi Shripad, Wels Christian, Beham-Schmid Christine, Fukunaga-Kalabis Mizuho, Holmen Sheri L, Otte Marcus, Herlyn Meenhard, Waldhoer Maria, Schaider Helmut
Cancer Biology Unit, Department of Dermatology, Medical University of Graz, Graz, Austria.
Centre for Medical Research (ZMF), Medical University of Graz, Graz, Austria.
Int J Cancer. 2015 Sep 15;137(6):1503-8. doi: 10.1002/ijc.29506. Epub 2015 Mar 16.
US28, a constitutively active G-protein-coupled receptor encoded by the human cytomegalovirus, leads to mechanistically unknown programmed cell death. Here we show that expression of wild-type US28 in human melanoma cells leads to apoptotic cell death via caspase 3 activation along with reduced cell proliferation. Reduced tumor growth upon US28 expression was observed in a xenograft mouse model. The signaling mute US28R129A showed a reduced antiproliferative effect. On evaluating different G-proteins coupled to US28 for signal transduction, Gα13 was identified as the main G-protein executing the apoptotic effect. Silencing of Gα13 but not Gαq resulted in a substantial increase in cell survival. Overexpression of Gα13 but not Gαq and their GTPase deficient forms Gα13Q226L and GαqQ209L, respectively, confirmed the requirement of Gα13 for US28 mediated cell death. Increasing expression of Gα13 alone induced cell death underscoring its relay function for US28 mediated decreased cell viability. Further reduced expression of Gα13 in melanoma cell lines isolated from advanced lesions and melanoma tissue was observed. These findings identified Gα13 as crucial for US28-induced cell death, substantiating that the effect of US28 on cell fate depends on preferred G-protein binding.
人巨细胞病毒编码的组成型活性G蛋白偶联受体US28会导致机制不明的程序性细胞死亡。我们在此表明,野生型US28在人黑色素瘤细胞中的表达会通过激活半胱天冬酶3导致凋亡性细胞死亡,同时细胞增殖减少。在异种移植小鼠模型中观察到US28表达后肿瘤生长减缓。信号传导沉默的US28R129A显示出降低的抗增殖作用。在评估与US28偶联以进行信号转导的不同G蛋白时,Gα13被确定为执行凋亡作用的主要G蛋白。沉默Gα13而非Gαq会导致细胞存活率大幅提高。分别过表达Gα13而非Gαq及其GTP酶缺陷形式Gα13Q226L和GαqQ209L,证实了Gα13对US28介导的细胞死亡的必要性。单独增加Gα13的表达会诱导细胞死亡,突出了其对US28介导的细胞活力降低的中继功能。在从晚期病变和黑色素瘤组织分离的黑色素瘤细胞系中观察到Gα13的表达进一步降低。这些发现确定Gα13对US28诱导的细胞死亡至关重要,证实了US28对细胞命运的影响取决于优先的G蛋白结合。