Direct and biologically significant interactions of human herpesvirus 8 interferon regulatory factor 1 with STAT3 and Janus kinase TYK2.
Direct and biologically significant interactions of human herpesvirus 8 interferon regulatory factor 1 with STAT3 and Janus kinase TYK2.
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DOI：
10.1371/journal.ppat.1011806
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发表时间：
2023-11
期刊：
影响因子：
6.7
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中科院分区：
文献类型：
--
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人类疱疹病毒 8 (HHV-8) 编码四种病毒干扰素调节因子 (vIRF)，这些因子以细胞 IRF 和/或其他先天免疫和应激信号调节因子为目标，并抑制细胞对病毒感染和复制的反应。对于 vIRF-1，细胞蛋白靶点包括 IRF、p53、p53 激活 ATM 激酶、BH3-only 蛋白以及抗病毒信号效应器 MAVS 和 STING； vIRF-1 抑制每一种，并已证实或可能促进 HHV-8 从头感染和有效复制。在这里，我们确定了 vIRF-1 与 STAT3 和 STAT 激活 Janus 激酶 TYK2（我们之前报道后者被 vIRF-1 抑制）的直接相互作用以及 vIRF-1 对细胞因子诱导的 STAT3 激活的抑制。在转染细胞中，vIRF-1 对活性磷酸化 STAT3 (pSTAT3) 的抑制是明显的，而在裂解性再激活的重组 HHV-8 感染细胞中，vIRF-1 消除导致 pSTAT3 水平升高。使用一组 vIRF-1 缺失变体，鉴定了与 STAT3 和 TYK2 相互作用所需的 vIRF-1 区域，这使得 vIRF-1 的 STAT3 信号传导抑制与 TYK2 结合相关联，而与 STAT3 相互作用无关。表达 vIRF-1 缺失变体 Δ198-222 的病毒突变体对 TYK2 相互作用和 pSTAT3 抑制无效，其有效复制受到严重损害。相反，磷酸酶抗性、长期活性的 STAT3 的表达会导致 HHV-8 复制受损。感染表达 STAT3 难治性 vIRF-1 缺失变体或 STAT3 耗尽的 HHV-8 突变体的细胞显示出 vIRF-1 表达降低，而定制肽促进的 STAT3 相互作用可能会增加 vIRF-1 表达并增强病毒复制。综上所述，我们的数据确定了 vIRF-1 靶向和 TYK2 抑制是 STAT3 信号传导抑制的机制，对 HHV-8 生产性复制至关重要，特定 pSTAT3 水平对复制的重要性，STAT3 和 vIRF-1-STAT3 相互作用在 vIRF-1 表达中的积极作用，以及 vIRF-1 对 STAT3 靶向裂解复制的重要贡献。 HHV-8 在病因学上与卡波西肉瘤、原发性渗出性淋巴瘤 (PEL) 和多中心卡斯尔曼病有关，这些疾病主要发生在 HIV 合并感染的情况下。这些疾病缺乏有效的治疗方法，其中涉及 HHV-8 感染的潜伏模式和溶解模式。了解 HHV-8 的分子生物学以及对病毒产生和潜伏感染至关重要的病毒-细胞相互作用对于开发新型抗病毒和治疗策略非常重要。 HHV-8 vIRF-1 对于潜伏和裂解复制都具有重要意义，有助于潜伏感染的 PEL 细胞的活力，并且是高效病毒生产所必需的。这项研究确定了 vIRF-1 与信号蛋白 STAT3 和 STAT 激活 TYK2 激酶的直接相互作用，并将各自的相互作用与 vIRF-1 表达和受感染细胞中 STAT3 激活的抑制以及促进生产性复制相关联。这些数据为开发针对这些新发现的直接相互作用的新型特异性抗病毒药物奠定了基础。
Human herpesvirus 8 (HHV-8) encodes four viral interferon regulatory factors (vIRFs) that target cellular IRFs and/or other innate-immune and stress signaling regulators and suppress the cellular response to viral infection and replication. For vIRF-1, cellular protein targets include IRFs, p53, p53-activating ATM kinase, BH3-only proteins, and antiviral signaling effectors MAVS and STING; vIRF-1 inhibits each, with demonstrated or likely promotion of HHV-8 de novo infection and productive replication. Here, we identify direct interactions of vIRF-1 with STAT3 and STAT-activating Janus kinase TYK2 (the latter reported previously by us to be inhibited by vIRF-1) and suppression by vIRF-1 of cytokine-induced STAT3 activation. Suppression of active, phosphorylated STAT3 (pSTAT3) by vIRF-1 was evident in transfected cells and vIRF-1 ablation in lytically-reactivated recombinant-HHV-8-infected cells led to increased levels of pSTAT3. Using a panel of vIRF-1 deletion variants, regions of vIRF-1 required for interactions with STAT3 and TYK2 were identified, which enabled correlation of STAT3 signaling inhibition by vIRF-1 with TYK2 binding, independently of STAT3 interaction. A viral mutant expressing vIRF-1 deletion-variant Δ198–222 refractory for TYK2 interaction and pSTAT3 suppression was severely compromised for productive replication. Conversely, expression of phosphatase-resistant, protractedly-active STAT3 led to impaired HHV-8 replication. Cells infected with HHV-8 mutants expressing STAT3-refractory vIRF-1 deletion variants or depleted of STAT3 displayed reduced vIRF-1 expression, while custom-peptide-promoted STAT3 interaction could effect increased vIRF-1 expression and enhanced virus replication. Taken together, our data identify vIRF-1 targeting and inhibition of TYK2 as a mechanism of STAT3-signaling suppression and critical for HHV-8 productive replication, the importance of specific pSTAT3 levels for replication, positive roles of STAT3 and vIRF-1-STAT3 interaction in vIRF-1 expression, and significant contributions to lytic replication of STAT3 targeting by vIRF-1. HHV-8 is involved etiologically in Kaposi’s sarcoma, primary effusion lymphoma (PEL), and multicentric Castleman’s disease, which occur predominantly in the context of HIV co-infection. Effective treatments for these diseases, in which both latent and lytic modes of HHV-8 infection are implicated, are lacking. Understanding the molecular biology of HHV-8 and the virus-cellular interactions critical for virus productive and latent infection is important for the development of novel antiviral and therapeutic strategies. HHV-8 vIRF-1 is of demonstrated importance to both latency and lytic replication, contributing to latently-infected PEL cell viability and being required for efficient virus production. This study identifies direct interactions of vIRF-1 with signaling protein STAT3 and STAT-activating TYK2 kinase and correlates the respective interactions with vIRF-1 expression and suppression of STAT3 activation in infected cells along with promotion of productive replication. These data provide a foundation for the development of novel and specific antiviral agents targeting these newly-identified direct interactions.
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影响因子：
3.7
作者：
Chinnakannan SK;Nanda SK;Baron MD
通讯作者：
Baron MD
DOI：
10.1073/pnas.171311298
发表时间：
2001-08-14
影响因子：
11.1
作者：
Gong, GZ;Waris, G;Siddiqui, A
通讯作者：
Siddiqui, A
影响因子：
5.4
作者：
Chen, Daming;Sandford, Gordon;Nicholas, John
通讯作者：
Nicholas, John
影响因子：
8
作者：
Gao, SJ;Boshoff, C;Moore, PS
通讯作者：
Moore, PS
影响因子：
56.9
作者：
Chatterjee, M;Osborne, J;Moore, PS
通讯作者：
Moore, PS