Herpes simplex virus type IICP27-Dependent activation of NF-κB
Herpes simplex virus type IICP27-Dependent activation of NF-κB
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DOI：
10.1128/jvi.01119-06
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发表时间：
2006-11-01
影响因子：
5.4
通讯作者：
Bachenheimer, Steven L.
中科院分区：
文献类型：
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作者：
Hargett, Danna;Rice, Stephen;Bachenheimer, Steven L.
单纯疱疹病毒1型（HSV-1）激活NF-κ B的能力已被充分证明。从感染后3至5小时开始，HSV-1诱导NF-κ B依赖性（p50/p65异源二聚体）DNA结合活性的稳定和持续的核转位，通过电泳迁移率变动测定。激活需要病毒结合和进入，以及从头感染细胞蛋白质合成，并伴随着I κ B α和I κ B β的丢失。在这项研究中，我们确定了I κ B α的丢失作为NF-κ B激活的标志物，并且用缺失个别立即早期（IE）调节蛋白的突变体感染表明ICP 27是I κ B α丢失所必需的。对ICP 27的N-末端和C-末端突变体的分析鉴定了从氨基酸21至63的区域为I κ B α丢失所必需的。用IE基因缺失的突变体病毒进行的其他实验显示，NF-κ B活化的ICP 27依赖性机制可能被功能性ICP 4增强。我们还分析了NF-κ B活化的另外两个标志物，即丝氨酸276和丝氨酸536上p65亚基的磷酸化。两种丝氨酸的磷酸化在HSV感染后被诱导，并且需要功能性ICP 4和ICP 27。药理学抑制剂研究表明，I κ B α和Ser 276磷酸化依赖于Jun N-末端蛋白激酶活性，而Ser 536磷酸化在抑制剂治疗期间不受影响。这些结果表明，有几个层次的调节NF-κ B激活在HSV感染，突出的重要作用，NF-κ B可能在感染中发挥。
The ability of herpes simplex virus type 1 (HSV-1) to activate NF-kappa B has been well documented. Beginning at 3 to 5 h postinfection, HSV-1 induces a robust and persistent nuclear translocation of an NF-kappa B-dependent (p50/p65 heterodimer) DNA binding activity, as measured by electrophoretic mobility shift assay. Activation requires virus binding and entry, as well as de novo infected-cell protein synthesis, and is accompanied by loss of both I kappa B alpha and I kappa B beta. In this study, we identified loss of I kappa B alpha as a marker of NF-kappa B activation, and infection with mutants with individual immediate-early (IE) regulatory proteins deleted indicated that ICP27 was necessary for I kappa B alpha loss. Analysis of both N-terminal and C-terminal mutants of ICP27 identified the region from amino acids 21 to 63 as being necessary for I kappa B alpha loss. Additional experiments with mutant viruses with combinations of IE genes deleted revealed that the ICP27-dependent mechanism of NF-kappa B activation may be augmented by functional ICP4. We also analyzed two additional markers for NF-kappa B activation, phosphorylation of the p65 subunit on Ser276 and Ser536. Phosphorylation of both serines was induced upon HSV infection and required functional ICP4 and ICP27. Pharmacological inhibitor studies revealed that both I kappa B alpha and Ser276 phosphorylation were dependent on Jun N-terminal protein kinase activity, while Ser536 phosphorylation was not affected during inhibitor treatment. These results demonstrate that there are several layers of regulation of NF-kappa B activation during HSV infection, highlighting the important role that NF-kappa B may play in infection.