Xuan Baoqin, Qian Zhikang, Torigoi Emi, Yu Dong
Department of Molecular Microbiology, Washington University School of Medicine, St. Louis, MO 63110, USA.
J Virol. 2009 Apr;83(8):3463-74. doi: 10.1128/JVI.02307-08. Epub 2009 Feb 4.
The endoplasmic reticulum (ER) is a key organelle involved in sensing and responding to stressful conditions, including those resulting from infection of viruses, such as human cytomegalovirus (HCMV). Three signaling pathways collectively termed the unfolded protein response (UPR) are activated to resolve ER stress, but they will also lead to cell death if the stress cannot be alleviated. HCMV is able to modulate the UPR to promote its infection. The specific viral factors involved in such HCMV-mediated modulation, however, were unknown. We previously showed that HCMV protein pUL38 was required to maintain the viability of infected cells, and it blocked cell death induced by thapsigargin. Here, we report that pUL38 is an HCMV-encoded regulator to modulate the UPR. In infection, pUL38 allowed HCMV to upregulate phosphorylation of PKR-like ER kinase (PERK) and the alpha subunit of eukaryotic initiation factor 2 (eIF-2alpha), as well as induce robust accumulation of activating transcriptional factor 4 (ATF4), key components of the PERK pathway. pUL38 also allowed the virus to suppress persistent phosphorylation of c-Jun N-terminal kinase (JNK), which was induced by the inositol-requiring enzyme 1 pathway. In isolation, pUL38 overexpression elevated eIF-2alpha phosphorylation, induced ATF4 accumulation, limited JNK phosphorylation, and suppressed cell death induced by both thapsigargin and tunicamycin, two drugs that induce ER stress by different mechanisms. Importantly, ATF4 overexpression and JNK inhibition significantly reduced cell death in pUL38-deficient virus infection. Thus, pUL38 targets ATF4 expression and JNK activation, and this activity appears to be critical for protecting cells from ER stress induced by HCMV infection.
内质网(ER)是一种关键细胞器,参与感知和应对应激条件,包括由病毒感染(如人类巨细胞病毒(HCMV))导致的应激。三种统称为未折叠蛋白反应(UPR)的信号通路被激活以解决内质网应激,但如果应激无法缓解,它们也会导致细胞死亡。HCMV能够调节UPR以促进其感染。然而,参与这种HCMV介导调节的具体病毒因子尚不清楚。我们之前表明,HCMV蛋白pUL38是维持感染细胞活力所必需的,并且它能阻止毒胡萝卜素诱导的细胞死亡。在此,我们报道pUL38是一种由HCMV编码的调节UPR的因子。在感染过程中,pUL38使HCMV能够上调蛋白激酶R样内质网激酶(PERK)和真核起始因子2(eIF-2α)的α亚基的磷酸化,以及诱导激活转录因子4(ATF4)的大量积累,ATF4是PERK通路的关键组成部分。pUL38还使病毒能够抑制由肌醇需求酶1通路诱导的c-Jun氨基末端激酶(JNK)的持续磷酸化。单独来看,pUL38的过表达提高了eIF-2α的磷酸化,诱导了ATF4的积累,限制了JNK的磷酸化,并抑制了由毒胡萝卜素和衣霉素诱导的细胞死亡,这两种药物通过不同机制诱导内质网应激。重要的是,ATF4的过表达和JNK的抑制显著降低了pUL38缺陷病毒感染中的细胞死亡。因此,pUL38靶向ATF4的表达和JNK的激活,并且这种活性似乎对于保护细胞免受HCMV感染诱导的内质网应激至关重要。