Dysregulation of Cellular VRK1, BAF, and Innate Immune Signaling by the Vaccinia Virus B12 Pseudokinase.
Dysregulation of Cellular VRK1, BAF, and Innate Immune Signaling by the Vaccinia Virus B12 Pseudokinase.
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DOI：
10.1128/jvi.00398-22
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发表时间：
2022-06-08
影响因子：
5.4
通讯作者：
中科院分区：
文献类型：
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痘病毒蛋白通过靶向宿主酶进行抑制和重定向来重塑整个细胞的信号传导。最近，人们发现，在感染早期，痘苗病毒 (VACV) B12 假激酶与细胞核中的细胞激酶 VRK1（一种前病毒因子）共纯化。尽管这种复合物的形成与细胞质 VACV DNA 复制的抑制相关，并且可能具有其他下游信号传导后果，但所涉及的分子机制仍知之甚少。在这里，我们进一步描述了 B12 和 VRK1 在痘病毒感染过程中如何相互调节。首先，我们证明 B12 在 VRK1 存在的情况下稳定，并且 VRK1 和 B12 共同影响它们各自的溶解度和亚细胞定位。在这方面，我们发现 B12 在有丝分裂过程中促进 VRK1 与细胞 DNA 共定位，并且 B12 和 VRK1 可能协同束缚于染色质。接下来，我们观察到 VRK1 的 C 末端尾部对于 B12-VRK1 复合物的形成或其前病毒活性来说是不必要的。有趣的是，我们发现了 B12 的点突变，能够消除与 VRK1 的相互作用，并使 B12 在感染过程中不受抑制。最后，我们研究了B12对宿主因子BAF和抗病毒信号通路的影响，发现B12触发BAF从细胞质到细胞核的重新分配。此外，B12 还能增强 DNA 诱导的先天免疫信号传导，揭示 B12 假激酶的新功能结果。总之，这项研究描述了 B12 在痘病毒感染过程中发挥的多方面作用，影响 VRK1、BAF 和先天免疫信号。重要性 蛋白质假激酶占人类激酶组以及其他生命形式的相当一部分。最近的研究表明，与激酶对应物相比，它们缺乏关键催化残基，但这并不能否定它们与分子信号转导相交叉的能力。虽然假激酶可以发挥多方面的作用，但它们对病毒感染的贡献仍未得到充分研究。在这里，我们进一步表征了痘病毒感染期间 VACV B12 假激酶和人 VRK1 激酶如何在细胞核中相互调节并抑制 VACV DNA 复制的机制。我们发现 B12 破坏了 VRK1 及其下游靶标 BAF 的调节，同时还增强了 DNA 依赖性先天免疫信号传导。结合之前的数据，这些研究有助于痘病毒靶向核通路领域的不断发展，并提供 B12 在激活抗病毒免疫中尚未探索的作用的证据。
Poxvirus proteins remodel signaling throughout the cell by targeting host enzymes for inhibition and redirection. Recently, it was discovered that early in infection the vaccinia virus (VACV) B12 pseudokinase copurifies with the cellular kinase VRK1, a proviral factor, in the nucleus. Although the formation of this complex correlates with inhibition of cytoplasmic VACV DNA replication and likely has other downstream signaling consequences, the molecular mechanisms involved are poorly understood. Here, we further characterize how B12 and VRK1 regulate one another during poxvirus infection. First, we demonstrate that B12 is stabilized in the presence of VRK1 and that VRK1 and B12 coinfluence their respective solubility and subcellular localization. In this regard, we find that B12 promotes VRK1 colocalization with cellular DNA during mitosis and that B12 and VRK1 may be tethered cooperatively to chromatin. Next, we observe that the C-terminal tail of VRK1 is unnecessary for B12-VRK1 complex formation or its proviral activity. Interestingly, we identify a point mutation of B12 capable of abrogating interaction with VRK1 and which renders B12 nonrepressive during infection. Lastly, we investigated the influence of B12 on the host factor BAF and antiviral signaling pathways and find that B12 triggers redistribution of BAF from the cytoplasm to the nucleus. In addition, B12 increases DNA-induced innate immune signaling, revealing a new functional consequence of the B12 pseudokinase. Together, this study characterizes the multifaceted roles B12 plays during poxvirus infection that impact VRK1, BAF, and innate immune signaling. IMPORTANCE Protein pseudokinases comprise a considerable fraction of the human kinome, as well as other forms of life. Recent studies have demonstrated that their lack of key catalytic residues compared to their kinase counterparts does not negate their ability to intersect with molecular signal transduction. While the multifaceted roles pseudokinases can play are known, their contribution to virus infection remains understudied. Here, we further characterize the mechanism of how the VACV B12 pseudokinase and human VRK1 kinase regulate one another in the nucleus during poxvirus infection and inhibit VACV DNA replication. We find that B12 disrupts regulation of VRK1 and its downstream target BAF, while also enhancing DNA-dependent innate immune signaling. Combined with previous data, these studies contribute to the growing field of nuclear pathways targeted by poxviruses and provide evidence of unexplored roles of B12 in the activation of antiviral immunity.
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DOI：
10.1073/pnas.89.11.4825
发表时间：
1992-06-01
影响因子：
11.1
作者：
CHANG, HW;WATSON, JC;JACOBS, BL
通讯作者：
JACOBS, BL
DOI：
10.1083/jcb.153.4.865
发表时间：
2001-05-14
期刊：
The Journal of cell biology
影响因子：
--
作者：
Adams RR;Maiato H;Earnshaw WC;Carmena M
通讯作者：
Carmena M
影响因子：
64.8
作者：
Avgousti DC;Herrmann C;Kulej K;Pancholi NJ;Sekulic N;Petrescu J;Molden RC;Blumenthal D;Paris AJ;Reyes ED;Ostapchuk P;Hearing P;Seeholzer SH;Worthen GS;Black BE;Garcia BA;Weitzman MD
通讯作者：
Weitzman MD
DOI：
10.1038/s41577-019-0131-x
发表时间：
2019-04
期刊：
Nature reviews. Immunology
影响因子：
--
作者：
Altan-Bonnet G;Mukherjee R
通讯作者：
Mukherjee R
DOI：
10.1038/2345
发表时间：
1998-10-01
期刊：
NATURE STRUCTURAL BIOLOGY
影响因子：
--
作者：
Cai, M;Huang, Y;Clore, GM
通讯作者：
Clore, GM