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本研究通过阐明MEK1和MEK2亚型在单纯疱疹病毒Ⅱ型(herpes simplex virus type 2,HSV2)复制中介导的Raf/MEK/ERK(简称ERK)通路活化中的作用,以期进一步阐明该通路调控病毒复制的机制.研究中应用了MEK抑制剂U0126、针对MEK1和MEK2的特异性小干扰RNA(small interfering RNA,siRNA),以及用死、活病毒攻击易感细胞,分别检测细胞ERK通路的激活和病毒复制的水平.结果表明,HSV2活病毒感染能引起ERK通路双相激活,U0126则能有效地抑制该通路的活化以及病毒复制;而死病毒只能引起ERK通路早期时相激活;单独敲降(knockdown)MEK1可抑制死、活病毒引发的ERK通路早期时相激活及活病毒引发的晚期时相激活,而敲降MEK2未见对病毒引起的该通路双相激活产生显著影响.提示HSV2复制引起ERK通路双相激活是基于MEK1-ERKs方式调控的,这种信号蛋白激活传递模式在HSV2整个复制周期中具有重要的正调控作用.该结果进一步证明了本室的前期报道:MEK1和MEK2亚型在病毒复制时发挥的作用不同.这对揭示MEK激酶功能的复杂性和ERK通路调控HSV2复制的机制,具有重要的意义.
This study was designed to elucidate the role of MEK1 and MEK2 subtypes in the activation of Raf / MEK / ERK (ERK) pathway mediated by herpes simplex virus type 2 (HSV2) replication in order to further elucidate the pathway regulation The mechanism of virus replication was studied.The MEK inhibitor U0126, small interfering RNA (siRNA) targeting MEK1 and MEK2, and the activation of ERK signaling pathway And viral replication level.The results showed that HSV2 virus infection can cause biphasic activation of ERK pathway, U0126 can effectively inhibit the activation of the pathway and virus replication; and the death virus can only cause ERK pathway early phase activation; MEK1 knockdown inhibited the early phase activation of dead and live virus-induced ERK pathway and the activation of live virus-induced late phase, while knockdown of MEK2 showed no significant effect on the biphasic activation of this pathway caused by the virus, suggesting that HSV2 Biphasic activation of ERK pathway by replication was regulated by the MEK1-ERKs mode, which has an important positive regulative effect during the entire replication cycle of HSV2. Previous reports demonstrated early this room: MEK1 and MEK2 isoforms play a role in viral replication This is different to reveal the complexity of the MEK kinase function and regulation mechanism HSV2 copy ERK pathway has important significance.