Department of Neurology/MC2030, The University of Chicago, 5841 S. Maryland Ave., Chicago, IL 60637, USA.
J Virol. 2011 Jul;85(14):7177-85. doi: 10.1128/JVI.00009-11. Epub 2011 May 11.
Cellular apoptosis induced by viral genes can play a critical role in determining virulence as well as viral persistence. This form of cell death has been of interest with respect to Theiler's murine encephalomyelitis virus (TMEV) because the GDVII strain and members of the GDVII subgroup are highly neurovirulent, while the DA strain and members of the TO subgroup induce a chronic progressive inflammatory demyelination with persistence of the virus in the central nervous system. The TMEV L protein has been identified as important in the pathogenesis of Theiler's virus-induced demyelinating disease (TMEV-IDD). We now show that DA L is apoptotic following transfection of L expression constructs or following DA virus infection of HeLa cells; the apoptotic activity depends on the presence of the serine/threonine domain of L, especially a serine at amino acid 57. In contrast, GDVII L has little apoptotic activity following transfection of L expression constructs in HeLa cells and is antiapoptotic following GDVII infection of HeLa cells. Of note, both DA and GDVII L cleave caspase-3 in BHK-21 cells, although neither implements the full apoptotic machinery in this cell type as manifested by the induction of terminal deoxynucleotidyltransferase-mediated dUTP-biotin nick end labeling (TUNEL) staining. The differences in apoptotic activities of DA and GDVII L in varied cell types may play an important role in TMEV subgroup-specific disease phenotypes.
病毒基因诱导的细胞凋亡在决定毒力和病毒持续存在方面起着至关重要的作用。这种形式的细胞死亡一直是 Theiler 鼠脑脊髓炎病毒 (TMEV) 关注的焦点,因为 GDVII 株和 GDVII 亚组的成员具有高度神经毒性,而 DA 株和 TO 亚组的成员则诱导慢性进行性炎症性脱髓鞘,病毒在中枢神经系统中持续存在。TMEV L 蛋白已被确定在 Theiler 病毒诱导的脱髓鞘病 (TMEV-IDD) 的发病机制中很重要。我们现在表明,DA L 在转染 L 表达构建体或 DA 病毒感染 HeLa 细胞后会发生凋亡;凋亡活性取决于 L 的丝氨酸/苏氨酸结构域的存在,特别是 57 位氨基酸的丝氨酸。相比之下,在 HeLa 细胞中转染 L 表达构建体后,GDVII L 的凋亡活性很小,而在 HeLa 细胞中感染 GDVII 后则具有抗凋亡活性。值得注意的是,DA 和 GDVII L 均可在 BHK-21 细胞中切割半胱天冬酶-3,尽管在这种细胞类型中,两者都没有实施完整的凋亡机制,如末端脱氧核苷酸转移酶介导的 dUTP-生物素 nick 末端标记 (TUNEL) 染色诱导所证明的那样。在不同细胞类型中 DA 和 GDVII L 的凋亡活性差异可能在 TMEV 亚组特异性疾病表型中发挥重要作用。