Vimentin rearrangement during African swine fever virus infection involves retrograde transport along microtubules and phosphorylation of vimentin by calcium calmodulin kinase II
Vimentin rearrangement during African swine fever virus infection involves retrograde transport along microtubules and phosphorylation of vimentin by calcium calmodulin kinase II
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DOI：
10.1128/jvi.79.18.11766-11775.2005
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发表时间：
2005-09-01
影响因子：
5.4
通讯作者：
Wileman, T
中科院分区：
文献类型：
--
作者：
Stefanovic, S;Windsor, M;Wileman, T
非洲猪瘟病毒（ASFV）感染导致弧菌蛋白重新排列到病毒工厂周围的笼子中。蛋白在细胞中的重排通常涉及细胞激酶对蛋白n端结构域的磷酸化，以促进微管上蛋白丝的拆卸和运输。在这里，我们证明了在ASFV感染期间，静脉蛋白重排的第一阶段涉及到微管依赖浓度的静脉蛋白进入靠近微管组织中心的病毒组装位点的“aster”。在工厂形成的早期，aster可能起着结构性的作用。将紫菀转化成笼子需要ASFV DNA复制。有趣的是，病毒DNA复制还导致钙钙调素依赖性蛋白激酶II （CaM激酶II）的激活和丝氨酸82上vimentin n端结构域的磷酸化。免疫染色显示笼内vimentin在丝氨酸82上被磷酸化。值得注意的是，病毒DNA复制和Ser 82磷酸化都被CaM激酶II的抑制剂KN93阻断，这表明CaM激酶II的激活、DNA复制和晚期基因表达之间存在联系。vimentin在丝氨酸82上的磷酸化可能是笼形形成所必需的，也可能仅仅是ASFV激活CaM激酶II的结果。蛋白笼可能具有细胞保护功能，防止病毒成分进入细胞质，同时在病毒组装位点集中晚期结构蛋白。
African swine fever virus (ASFV) infection leads to rearrangement of vimentin into a cage surrounding virus factories. Vimentin rearrangement in cells generally involves phosphorylation of N-terminal domains of vimentin by cellular kinases to facilitate disassembly and transport of vimentin filaments on microtubules. Here, we demonstrate that the first stage in vimentin rearrangement during ASFV infection involves a microtubule-dependent concentration of vimentin into an "aster" within virus assembly sites located close to the microtubule organizing center. The aster may play a structural role early during the formation of the factory. Conversion of the aster into a cage required ASFV DNA replication. Interestingly, viral DNA replication also resulted in the activation of calcium calmodulin-dependent protein kinase II (CaM kinase II) and phosphorylation of the N-terminal domain of vimentin on serine 82. Immunostaining showed that vimentin within the cage was phosphorylated on serine 82. Significantly, both viral DNA replication and Ser 82 phosphorylation were blocked by KN93, an inhibitor of CaM kinase II, suggesting a link between CaM kinase II activation, DNA replication, and late gene expression. Phosphorylation of vimentin on serine 82 may be necessary for cage formation or may simply be a consequence of activation of CaM kinase II by ASFV. The vimentin cage may serve a cytoprotective function and prevent movement of viral components into the cytoplasm and at the same time concentrate late structural proteins at sites of virus assembly.