Department of Virology, Faculty of Medicine, University of Crete, Heraklion 71003, Crete, Greece.
Eur J Cell Biol. 2010 Oct;89(10):757-68. doi: 10.1016/j.ejcb.2010.05.006.
In controlling the switch from latency to lytic infection, the immediate early (IE) genes lie at the core of herpesvirus pathogenesis. To image the 72kDa human cytomegalovirus (HCMV) major IE protein (IE1-72K), a recombinant virus encoding IE1 fused with EGFP was constructed. Using this construct, the IE1-EGFP fusion was detected at ND10 (PML-bodies) within 2h post infection (p.i.) and the complete disruption of ND10 imaged through to 6h p.i. HCMV genomes and IE2-86K protein could be detected adjacent to the slowly degrading IE1-72K/ND10 foci. IE1-72K associates with metaphase chromatin, recruiting both PML and STAT2. hDaxx, STAT1 and IE2-86K did not re-locate to metaphase chromatin; the fate of hDaxx is particularly important as this protein contributes to an intrinsic barrier to HCMV infection. While IE1-72K participates in a complex with chromatin, PML, STAT2 and Sp100, IE1-72K releases hDaxx from ND10 yet does not appear to remain associated with it.
在控制潜伏期向裂解感染的转变中，即刻早期（IE）基因是疱疹病毒发病机制的核心。为了对 72kDa 人巨细胞病毒（HCMV）主要 IE 蛋白（IE1-72K）进行成像，构建了一种编码与 EGFP 融合的 IE1 的重组病毒。利用该构建体，在感染后 2 小时（p.i.）即可在 ND10（PML 体）中检测到 IE1-EGFP 融合，并且可以在 6 小时 p.i. 之前观察到 ND10 的完全破坏。可以在邻近缓慢降解的 IE1-72K/ND10 焦点处检测到 HCMV 基因组和 IE2-86K 蛋白。IE1-72K 与中期染色质结合，招募 PML 和 STAT2。hDaxx、STAT1 和 IE2-86K 没有重新定位到中期染色质；hDaxx 的命运尤为重要，因为该蛋白有助于对 HCMV 感染产生内在屏障。虽然 IE1-72K 与染色质、PML、STAT2 和 Sp100 形成复合物，但 IE1-72K 从 ND10 释放 hDaxx，但似乎不再与之相关。
