Division of Molecular Virology, Department of Microbiology and Immunology, The Institute of Medical Science, The University of Tokyo, Tokyo, Japan.
Department of Infectious Disease Control, International Research Center for Infectious Diseases, The Institute of Medical Science, The University of Tokyo, Tokyo, Japan.
J Virol. 2024 Aug 20;98(8):e0073724. doi: 10.1128/jvi.00737-24. Epub 2024 Jul 17.
Facilitates chromatin transcription (FACT) interacts with nucleosomes to promote gene transcription by regulating the dissociation and reassembly of nucleosomes downstream and upstream of RNA polymerase II (Pol II). A previous study reported that herpes simplex virus 1 (HSV-1) regulatory protein ICP22 interacted with FACT and was required for its recruitment to the viral DNA genome in HSV-1-infected cells. However, the biological importance of interactions between ICP22 and FACT in relation to HSV-1 infection is unclear. Here, we mapped the minimal domain of ICP22 required for its efficient interaction with FACT to a cluster of five basic amino acids in ICP22. A recombinant virus harboring alanine substitutions in this identified cluster led to the decreased accumulation of viral mRNAs from UL54, UL38, and UL44 genes, reduced Pol II occupancy of these genes in MRC-5 cells, and impaired HSV-1 virulence in mice following ocular or intracranial infection. Furthermore, the treatment of mice infected with wild-type HSV-1 with CBL0137, a FACT inhibitor currently being investigated in clinical trials, significantly improved the survival rate of mice. These results suggested that the interaction between ICP22 and FACT was required for efficient HSV-1 gene expression and pathogenicity. Therefore, FACT might be a potential therapeutic target for HSV-1 infection.IMPORTANCEICP22 is a well-known regulatory factor of HSV-1 gene expression, but its mechanism(s) are poorly understood. Although the interaction of FACT with ICP22 was reported previously, its significance in HSV-1 infection is unknown. Given that FACT is involved in gene transcription, it is of interest to investigate this interaction as it relates to HSV-1 gene expression. To determine a direct link between the interaction and HSV-1 infection, we mapped a minimal domain of ICP22 required for its efficient interaction with FACT and generated a recombinant virus carrying mutations in the identified domain. Using the recombinant virus, we obtained evidence suggesting that the interaction between ICP22 and FACT promoted Pol II transcription from HSV-1 genes and viral virulence in mice. In addition, CBL0137, an inhibitor of FACT, effectively protected mice from lethal HSV-1 infection, suggesting FACT might be a potential target for the development of novel anti-HSV drugs.
促进染色质转录 (FACT) 与核小体相互作用,通过调节 RNA 聚合酶 II (Pol II) 下游和上游核小体的解聚和组装,促进基因转录。先前的研究表明,单纯疱疹病毒 1 (HSV-1) 调节蛋白 ICP22 与 FACT 相互作用,并且在 HSV-1 感染细胞中需要其募集到病毒 DNA 基因组。然而,ICP22 与 FACT 之间的相互作用与 HSV-1 感染的生物学重要性尚不清楚。在这里,我们将 ICP22 与其与 FACT 有效相互作用所需的最小结构域映射到 ICP22 中五个碱性氨基酸簇上。携带该鉴定簇中氨基酸替换的重组病毒导致 UL54、UL38 和 UL44 基因的病毒 mRNA 积累减少,MRC-5 细胞中这些基因的 Pol II 占有率降低,以及在眼部或颅内感染后 HSV-1 毒力受损。此外,用目前正在临床试验中研究的 FACT 抑制剂 CBL0137 治疗感染野生型 HSV-1 的小鼠,显著提高了小鼠的存活率。这些结果表明,ICP22 与 FACT 之间的相互作用对于 HSV-1 基因表达和致病性是必需的。因此,FACT 可能是 HSV-1 感染的潜在治疗靶点。
重要性 ICP22 是 HSV-1 基因表达的一个众所周知的调节因子,但它的机制尚不清楚。尽管先前已经报道了 FACT 与 ICP22 的相互作用,但它在 HSV-1 感染中的意义尚不清楚。鉴于 FACT 参与基因转录,研究这种相互作用与 HSV-1 基因表达的关系很有意义。为了确定相互作用与 HSV-1 感染之间的直接联系,我们绘制了 ICP22 与其与 FACT 有效相互作用所需的最小结构域,并生成了携带鉴定结构域中突变的重组病毒。使用重组病毒,我们获得了证据表明,ICP22 与 FACT 之间的相互作用促进了 HSV-1 基因的 Pol II 转录和病毒在小鼠中的毒力。此外,FACT 的抑制剂 CBL0137 有效地保护了小鼠免受致命的 HSV-1 感染,这表明 FACT 可能是开发新型抗 HSV 药物的潜在靶点。